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Karl J Kramer - One of the best experts on this subject based on the ideXlab platform.
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tyrosine metabolism for insect cuticle pigmentation and Sclerotization
2016Co-Authors: Yasuyuki Arakane, Tsunaki Asano, Karl J KramerAbstract:Pigmentation or body color patterns in insects quite often differ not only between species but also in different stages of development and in different body regions of a single species. Body coloration plays physiologically and ecologically important roles as for instance in species recognition and communication, courtship/mate selection, mimicry, crypsis, warning, prey-predator/parasite interactions, and resistance to temperature, desiccation and absorbs or reflects harmful ultraviolet radiation. Many kinds of pigment molecules and structural colors contribute to the diversity of body coloration in insects. Recent studies have elucidated some of the genetic and molecular biological mechanisms underlying pigment biosynthesis. This chapter focuses on the pigments derived from the amino acid tyrosine. The tyrosine-mediated cuticle tanning pathway is responsible for production of melanins and other pigments derived from 3,4-dihydroxyphenylalanine (DOPA) and dopamine as well as from N-acyldopamines. The N-acylated dopamines, in addition, are oxidized by the phenoloxidase laccase 2 to form quinones and quinone methides, which then undergo cross-linking reactions with cuticular proteins (CPs) for cuticle Sclerotization. We review the regulation and functional importance and also the diversity of the genes involved in this pathway. The unique localization and cross-linking of specific CPs for morphology and ultrastructure of the exoskeleton are also discussed.
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model reactions for insect cuticle Sclerotization participation of amino groups in the cross linking of manduca sexta cuticle protein mscp36
Insect Biochemistry and Molecular Biology, 2010Co-Authors: Richard J Suderman, Neal T Dittmer, Karl J Kramer, Michael R KanostAbstract:Current theories of Sclerotization center on protein cross-linking and dehydration as major factors in the hardening and stability of the insect cuticle. Several studies have reported the identification of catecholamino acid adducts from sclerotizing cuticle involving histidine, lysine, and tyrosine, though there have been no reports of a catechol linked between two amino acid residues. Previously, we reported an in vitro model system for Sclerotization and observed that stable protein oligomers were formed, presumably through cross-links with oxidized catecholamines [Insect Biochem. Mol. Biol. (2006) 36, 353e365]. Using site-directed mutagenesis we created a mutant lacking histidine, rMsCP36(H65A/H178A), to investigate the possible involvement of the two histidine residues of MsCP36 in cross-linking. Surprisingly, this alteration had little ornoeffecton the formation of proteinoligomers as determined bySDS-PAGEanalysis. Blockingof the freeaminogroupsinlysylsidechainsandtheamino-terminus bysuccinylationdiminished, but did not eliminate, cross-linking of either rMsCP36 or rMsCP36(H65A/H178A). We also examined the possibility that cross-linking was due to intermolecular dityrosine linkages. Immunoblot analysis utilizing a monoclonal antibody known to recognize peptidyl dityrosine indicated that dityrosyl cross-links were present. Taken together, these results indicate that lysyl residues are important for the cross-linking of the cuticle protein rMsCP36, but that additional residues other than histidine can also contribute. 2010 Elsevier Ltd. All rights reserved.
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erratumerratum to model reactions for insect cuticle Sclerotization cross linking of recombinant cuticular proteins upon their laccase catalyzed oxidative conjugation with catechols insect biochemistry and molecular biology 36 2006 353 365
Insect Biochemistry and Molecular Biology, 2006Co-Authors: Richard J Suderman, Neal T Dittmer, Michael R Kanost, Karl J KramerAbstract:Erratum to ‘‘Model reactions for insect cuticle Sclerotization: Cross-linking of recombinant cuticular proteins upon their laccase-catalyzed oxidative conjugation with catechols’’ [Insect Biochemistry and Molecular Biology 36 (2006) 353–365] Richard J. Suderman, Neal T. Dittmer, Michael R. Kanost , Karl J. Kramer Department of Biochemistry, 103 Willard Hall, Kansas State University, Manhattan, KS 66506-3706, USA Grain Marketing and Production Research Center, Agricultural Research Service, US Department of Agriculture, Manhattan, KS 66506-2736, USA
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model reactions for insect cuticle Sclerotization cross linking of recombinant cuticular proteins upon their laccase catalyzed oxidative conjugation with catechols
Insect Biochemistry and Molecular Biology, 2006Co-Authors: Richard J Suderman, Neal T Dittmer, Michael R Kanost, Karl J KramerAbstract:The quinone-tanning hypothesis for insect cuticle Sclerotization proposes that N-acylcatecholamines are oxidized by a phenoloxidase to quinones and quinone methides, which serve as electrophilic cross-linking agents to form covalent cross-links between cuticular proteins. We investigated model reactions for protein cross-linking that occurs during insect cuticle Sclerotization using recombinant pupal cuticular proteins from the tobacco hornworm, Manduca sexta, fungal or recombinant hornworm laccase-type phenoloxidase, and the cross-linking agent precursor N-acylcatecholamines, N-b-alanydopamine (NBAD) or N-acetyldopamine (NADA). Recombinant M. sexta pupal cuticular proteins MsCP36, MsCP20, and MsCP27 were expressed and purified to near homogeneity. Polyclonal antisera to these recombinant proteins recognized the native proteins in crude pharate brown-colored pupal cuticle homogenates. Furthermore, antisera to MsCP36, which contains a type-1 Rebers and Riddiford (RR-1) consensus sequence, also recognized an immunoreactive protein in homogenates of larval head capsule exuviae, indicating the presence of an RR-1 cuticular protein in a very hard, sclerotized and nonpigmented cuticle. All three of the proteins formed small and large oligomers stable to boiling SDS treatment under reducing conditions after reaction with laccase and the N-acylcatecholamines. The optimal reaction conditions for MsCP36 polymerization were 0.3 mM MsCP36, 7.4 mM NBAD and 1.0 U/ml fungal laccase. Approximately 5–10% of the monomer reacted to yield insoluble oligomers and polymers during the reaction, and the monomer also became increasingly insoluble in SDS solution after reaction with the oxidized NBAD. When NADA was used instead of NBAD, less oligomer formation occurred, and most of the protein remained soluble. Radiolabeled NADA became covalently bound to the MsCP36 monomer and oligomers during cross-linking. Recombinant Manduca laccase (MsLac2) also catalyzed the polymerization of MsCP36. These results support the hypothesis that during Sclerotization, insect cuticular proteins are oxidatively conjugated with catechols, a posttranslational process termed catecholation, and then become crosslinked, forming oligomers and subsequently polymers. r 2006 Elsevier Ltd. All rights reserved.
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model reactions for insect cuticle Sclerotization cross linking of recombinant cuticular proteins upon their laccase catalyzed oxidative conjugation with catechols
Insect Biochemistry and Molecular Biology, 2006Co-Authors: Richard J Suderman, Neal T Dittmer, Michael R Kanost, Karl J KramerAbstract:The quinone-tanning hypothesis for insect cuticle Sclerotization proposes that N-acylcatecholamines are oxidized by a phenoloxidase to quinones and quinone methides, which serve as electrophilic cross-linking agents to form covalent cross-links between cuticular proteins. We investigated model reactions for protein cross-linking that occurs during insect cuticle Sclerotization using recombinant pupal cuticular proteins from the tobacco hornworm, Manduca sexta, fungal or recombinant hornworm laccase-type phenoloxidase, and the cross-linking agent precursor N-acylcatecholamines, N-b-alanydopamine (NBAD) or N-acetyldopamine (NADA). Recombinant M. sexta pupal cuticular proteins MsCP36, MsCP20, and MsCP27 were expressed and purified to near homogeneity. Polyclonal antisera to these recombinant proteins recognized the native proteins in crude pharate brown-colored pupal cuticle homogenates. Furthermore, antisera to MsCP36, which contains a type-1 Rebers and Riddiford (RR-1) consensus sequence, also recognized an immunoreactive protein in homogenates of larval head capsule exuviae, indicating the presence of an RR-1 cuticular protein in a very hard, sclerotized and nonpigmented cuticle. All three of the proteins formed small and large oligomers stable to boiling SDS treatment under reducing conditions after reaction with laccase and the N-acylcatecholamines. The optimal reaction conditions for MsCP36 polymerization were 0.3 mM MsCP36, 7.4 mM NBAD and 1.0 U/ml fungal laccase. Approximately 5–10% of the monomer reacted to yield insoluble oligomers and polymers during the reaction, and the monomer also became increasingly insoluble in SDS solution after reaction with the oxidized NBAD. When NADA was used instead of NBAD, less oligomer formation occurred, and most of the protein remained soluble. Radiolabeled NADA became covalently bound to the MsCP36 monomer and oligomers during cross-linking. Recombinant Manduca laccase (MsLac2) also catalyzed the polymerization of MsCP36. These results support the hypothesis that during Sclerotization, insect cuticular proteins are oxidatively conjugated with catechols, a posttranslational process termed catecholation, and then become crosslinked, forming oligomers and subsequently polymers. r 2006 Elsevier Ltd. All rights reserved.
Richard J Suderman - One of the best experts on this subject based on the ideXlab platform.
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model reactions for insect cuticle Sclerotization participation of amino groups in the cross linking of manduca sexta cuticle protein mscp36
Insect Biochemistry and Molecular Biology, 2010Co-Authors: Richard J Suderman, Neal T Dittmer, Karl J Kramer, Michael R KanostAbstract:Current theories of Sclerotization center on protein cross-linking and dehydration as major factors in the hardening and stability of the insect cuticle. Several studies have reported the identification of catecholamino acid adducts from sclerotizing cuticle involving histidine, lysine, and tyrosine, though there have been no reports of a catechol linked between two amino acid residues. Previously, we reported an in vitro model system for Sclerotization and observed that stable protein oligomers were formed, presumably through cross-links with oxidized catecholamines [Insect Biochem. Mol. Biol. (2006) 36, 353e365]. Using site-directed mutagenesis we created a mutant lacking histidine, rMsCP36(H65A/H178A), to investigate the possible involvement of the two histidine residues of MsCP36 in cross-linking. Surprisingly, this alteration had little ornoeffecton the formation of proteinoligomers as determined bySDS-PAGEanalysis. Blockingof the freeaminogroupsinlysylsidechainsandtheamino-terminus bysuccinylationdiminished, but did not eliminate, cross-linking of either rMsCP36 or rMsCP36(H65A/H178A). We also examined the possibility that cross-linking was due to intermolecular dityrosine linkages. Immunoblot analysis utilizing a monoclonal antibody known to recognize peptidyl dityrosine indicated that dityrosyl cross-links were present. Taken together, these results indicate that lysyl residues are important for the cross-linking of the cuticle protein rMsCP36, but that additional residues other than histidine can also contribute. 2010 Elsevier Ltd. All rights reserved.
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erratumerratum to model reactions for insect cuticle Sclerotization cross linking of recombinant cuticular proteins upon their laccase catalyzed oxidative conjugation with catechols insect biochemistry and molecular biology 36 2006 353 365
Insect Biochemistry and Molecular Biology, 2006Co-Authors: Richard J Suderman, Neal T Dittmer, Michael R Kanost, Karl J KramerAbstract:Erratum to ‘‘Model reactions for insect cuticle Sclerotization: Cross-linking of recombinant cuticular proteins upon their laccase-catalyzed oxidative conjugation with catechols’’ [Insect Biochemistry and Molecular Biology 36 (2006) 353–365] Richard J. Suderman, Neal T. Dittmer, Michael R. Kanost , Karl J. Kramer Department of Biochemistry, 103 Willard Hall, Kansas State University, Manhattan, KS 66506-3706, USA Grain Marketing and Production Research Center, Agricultural Research Service, US Department of Agriculture, Manhattan, KS 66506-2736, USA
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model reactions for insect cuticle Sclerotization cross linking of recombinant cuticular proteins upon their laccase catalyzed oxidative conjugation with catechols
Insect Biochemistry and Molecular Biology, 2006Co-Authors: Richard J Suderman, Neal T Dittmer, Michael R Kanost, Karl J KramerAbstract:The quinone-tanning hypothesis for insect cuticle Sclerotization proposes that N-acylcatecholamines are oxidized by a phenoloxidase to quinones and quinone methides, which serve as electrophilic cross-linking agents to form covalent cross-links between cuticular proteins. We investigated model reactions for protein cross-linking that occurs during insect cuticle Sclerotization using recombinant pupal cuticular proteins from the tobacco hornworm, Manduca sexta, fungal or recombinant hornworm laccase-type phenoloxidase, and the cross-linking agent precursor N-acylcatecholamines, N-b-alanydopamine (NBAD) or N-acetyldopamine (NADA). Recombinant M. sexta pupal cuticular proteins MsCP36, MsCP20, and MsCP27 were expressed and purified to near homogeneity. Polyclonal antisera to these recombinant proteins recognized the native proteins in crude pharate brown-colored pupal cuticle homogenates. Furthermore, antisera to MsCP36, which contains a type-1 Rebers and Riddiford (RR-1) consensus sequence, also recognized an immunoreactive protein in homogenates of larval head capsule exuviae, indicating the presence of an RR-1 cuticular protein in a very hard, sclerotized and nonpigmented cuticle. All three of the proteins formed small and large oligomers stable to boiling SDS treatment under reducing conditions after reaction with laccase and the N-acylcatecholamines. The optimal reaction conditions for MsCP36 polymerization were 0.3 mM MsCP36, 7.4 mM NBAD and 1.0 U/ml fungal laccase. Approximately 5–10% of the monomer reacted to yield insoluble oligomers and polymers during the reaction, and the monomer also became increasingly insoluble in SDS solution after reaction with the oxidized NBAD. When NADA was used instead of NBAD, less oligomer formation occurred, and most of the protein remained soluble. Radiolabeled NADA became covalently bound to the MsCP36 monomer and oligomers during cross-linking. Recombinant Manduca laccase (MsLac2) also catalyzed the polymerization of MsCP36. These results support the hypothesis that during Sclerotization, insect cuticular proteins are oxidatively conjugated with catechols, a posttranslational process termed catecholation, and then become crosslinked, forming oligomers and subsequently polymers. r 2006 Elsevier Ltd. All rights reserved.
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model reactions for insect cuticle Sclerotization cross linking of recombinant cuticular proteins upon their laccase catalyzed oxidative conjugation with catechols
Insect Biochemistry and Molecular Biology, 2006Co-Authors: Richard J Suderman, Neal T Dittmer, Michael R Kanost, Karl J KramerAbstract:The quinone-tanning hypothesis for insect cuticle Sclerotization proposes that N-acylcatecholamines are oxidized by a phenoloxidase to quinones and quinone methides, which serve as electrophilic cross-linking agents to form covalent cross-links between cuticular proteins. We investigated model reactions for protein cross-linking that occurs during insect cuticle Sclerotization using recombinant pupal cuticular proteins from the tobacco hornworm, Manduca sexta, fungal or recombinant hornworm laccase-type phenoloxidase, and the cross-linking agent precursor N-acylcatecholamines, N-b-alanydopamine (NBAD) or N-acetyldopamine (NADA). Recombinant M. sexta pupal cuticular proteins MsCP36, MsCP20, and MsCP27 were expressed and purified to near homogeneity. Polyclonal antisera to these recombinant proteins recognized the native proteins in crude pharate brown-colored pupal cuticle homogenates. Furthermore, antisera to MsCP36, which contains a type-1 Rebers and Riddiford (RR-1) consensus sequence, also recognized an immunoreactive protein in homogenates of larval head capsule exuviae, indicating the presence of an RR-1 cuticular protein in a very hard, sclerotized and nonpigmented cuticle. All three of the proteins formed small and large oligomers stable to boiling SDS treatment under reducing conditions after reaction with laccase and the N-acylcatecholamines. The optimal reaction conditions for MsCP36 polymerization were 0.3 mM MsCP36, 7.4 mM NBAD and 1.0 U/ml fungal laccase. Approximately 5–10% of the monomer reacted to yield insoluble oligomers and polymers during the reaction, and the monomer also became increasingly insoluble in SDS solution after reaction with the oxidized NBAD. When NADA was used instead of NBAD, less oligomer formation occurred, and most of the protein remained soluble. Radiolabeled NADA became covalently bound to the MsCP36 monomer and oligomers during cross-linking. Recombinant Manduca laccase (MsLac2) also catalyzed the polymerization of MsCP36. These results support the hypothesis that during Sclerotization, insect cuticular proteins are oxidatively conjugated with catechols, a posttranslational process termed catecholation, and then become crosslinked, forming oligomers and subsequently polymers. r 2006 Elsevier Ltd. All rights reserved.
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model reactions for insect cuticle Sclerotization cross linking of recombinant cuticular proteins upon their laccase catalyzed oxidative conjugation with catechols
Insect Biochemistry and Molecular Biology, 2006Co-Authors: Richard J Suderman, Neal T Dittmer, Michael R Kanost, Karl J KramerAbstract:The quinone-tanning hypothesis for insect cuticle Sclerotization proposes that N-acylcatecholamines are oxidized by a phenoloxidase to quinones and quinone methides, which serve as electrophilic cross-linking agents to form covalent cross-links between cuticular proteins. We investigated model reactions for protein cross-linking that occurs during insect cuticle Sclerotization using recombinant pupal cuticular proteins from the tobacco hornworm, Manduca sexta, fungal or recombinant hornworm laccase-type phenoloxidase, and the cross-linking agent precursor N-acylcatecholamines, N-beta-alanydopamine (NBAD) or N-acetyldopamine (NADA). Recombinant M. sexta pupal cuticular proteins MsCP36, MsCP20, and MsCP27 were expressed and purified to near homogeneity. Polyclonal antisera to these recombinant proteins recognized the native proteins in crude pharate brown-colored pupal cuticle homogenates. Furthermore, antisera to MsCP36, which contains a type-1 Rebers and Riddiford (RR-1) consensus sequence, also recognized an immunoreactive protein in homogenates of larval head capsule exuviae, indicating the presence of an RR-1 cuticular protein in a very hard, sclerotized and nonpigmented cuticle. All three of the proteins formed small and large oligomers stable to boiling SDS treatment under reducing conditions after reaction with laccase and the N-acylcatecholamines. The optimal reaction conditions for MsCP36 polymerization were 0.3mM MsCP36, 7.4mM NBAD and 1.0U/mul fungal laccase. Approximately 5-10% of the monomer reacted to yield insoluble oligomers and polymers during the reaction, and the monomer also became increasingly insoluble in SDS solution after reaction with the oxidized NBAD. When NADA was used instead of NBAD, less oligomer formation occurred, and most of the protein remained soluble. Radiolabeled NADA became covalently bound to the MsCP36 monomer and oligomers during cross-linking. Recombinant Manduca laccase (MsLac2) also catalyzed the polymerization of MsCP36. These results support the hypothesis that during Sclerotization, insect cuticular proteins are oxidatively conjugated with catechols, a posttranslational process termed catecholation, and then become cross-linked, forming oligomers and subsequently polymers.
Michael R Kanost - One of the best experts on this subject based on the ideXlab platform.
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model reactions for insect cuticle Sclerotization participation of amino groups in the cross linking of manduca sexta cuticle protein mscp36
Insect Biochemistry and Molecular Biology, 2010Co-Authors: Richard J Suderman, Neal T Dittmer, Karl J Kramer, Michael R KanostAbstract:Current theories of Sclerotization center on protein cross-linking and dehydration as major factors in the hardening and stability of the insect cuticle. Several studies have reported the identification of catecholamino acid adducts from sclerotizing cuticle involving histidine, lysine, and tyrosine, though there have been no reports of a catechol linked between two amino acid residues. Previously, we reported an in vitro model system for Sclerotization and observed that stable protein oligomers were formed, presumably through cross-links with oxidized catecholamines [Insect Biochem. Mol. Biol. (2006) 36, 353e365]. Using site-directed mutagenesis we created a mutant lacking histidine, rMsCP36(H65A/H178A), to investigate the possible involvement of the two histidine residues of MsCP36 in cross-linking. Surprisingly, this alteration had little ornoeffecton the formation of proteinoligomers as determined bySDS-PAGEanalysis. Blockingof the freeaminogroupsinlysylsidechainsandtheamino-terminus bysuccinylationdiminished, but did not eliminate, cross-linking of either rMsCP36 or rMsCP36(H65A/H178A). We also examined the possibility that cross-linking was due to intermolecular dityrosine linkages. Immunoblot analysis utilizing a monoclonal antibody known to recognize peptidyl dityrosine indicated that dityrosyl cross-links were present. Taken together, these results indicate that lysyl residues are important for the cross-linking of the cuticle protein rMsCP36, but that additional residues other than histidine can also contribute. 2010 Elsevier Ltd. All rights reserved.
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erratumerratum to model reactions for insect cuticle Sclerotization cross linking of recombinant cuticular proteins upon their laccase catalyzed oxidative conjugation with catechols insect biochemistry and molecular biology 36 2006 353 365
Insect Biochemistry and Molecular Biology, 2006Co-Authors: Richard J Suderman, Neal T Dittmer, Michael R Kanost, Karl J KramerAbstract:Erratum to ‘‘Model reactions for insect cuticle Sclerotization: Cross-linking of recombinant cuticular proteins upon their laccase-catalyzed oxidative conjugation with catechols’’ [Insect Biochemistry and Molecular Biology 36 (2006) 353–365] Richard J. Suderman, Neal T. Dittmer, Michael R. Kanost , Karl J. Kramer Department of Biochemistry, 103 Willard Hall, Kansas State University, Manhattan, KS 66506-3706, USA Grain Marketing and Production Research Center, Agricultural Research Service, US Department of Agriculture, Manhattan, KS 66506-2736, USA
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model reactions for insect cuticle Sclerotization cross linking of recombinant cuticular proteins upon their laccase catalyzed oxidative conjugation with catechols
Insect Biochemistry and Molecular Biology, 2006Co-Authors: Richard J Suderman, Neal T Dittmer, Michael R Kanost, Karl J KramerAbstract:The quinone-tanning hypothesis for insect cuticle Sclerotization proposes that N-acylcatecholamines are oxidized by a phenoloxidase to quinones and quinone methides, which serve as electrophilic cross-linking agents to form covalent cross-links between cuticular proteins. We investigated model reactions for protein cross-linking that occurs during insect cuticle Sclerotization using recombinant pupal cuticular proteins from the tobacco hornworm, Manduca sexta, fungal or recombinant hornworm laccase-type phenoloxidase, and the cross-linking agent precursor N-acylcatecholamines, N-b-alanydopamine (NBAD) or N-acetyldopamine (NADA). Recombinant M. sexta pupal cuticular proteins MsCP36, MsCP20, and MsCP27 were expressed and purified to near homogeneity. Polyclonal antisera to these recombinant proteins recognized the native proteins in crude pharate brown-colored pupal cuticle homogenates. Furthermore, antisera to MsCP36, which contains a type-1 Rebers and Riddiford (RR-1) consensus sequence, also recognized an immunoreactive protein in homogenates of larval head capsule exuviae, indicating the presence of an RR-1 cuticular protein in a very hard, sclerotized and nonpigmented cuticle. All three of the proteins formed small and large oligomers stable to boiling SDS treatment under reducing conditions after reaction with laccase and the N-acylcatecholamines. The optimal reaction conditions for MsCP36 polymerization were 0.3 mM MsCP36, 7.4 mM NBAD and 1.0 U/ml fungal laccase. Approximately 5–10% of the monomer reacted to yield insoluble oligomers and polymers during the reaction, and the monomer also became increasingly insoluble in SDS solution after reaction with the oxidized NBAD. When NADA was used instead of NBAD, less oligomer formation occurred, and most of the protein remained soluble. Radiolabeled NADA became covalently bound to the MsCP36 monomer and oligomers during cross-linking. Recombinant Manduca laccase (MsLac2) also catalyzed the polymerization of MsCP36. These results support the hypothesis that during Sclerotization, insect cuticular proteins are oxidatively conjugated with catechols, a posttranslational process termed catecholation, and then become crosslinked, forming oligomers and subsequently polymers. r 2006 Elsevier Ltd. All rights reserved.
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model reactions for insect cuticle Sclerotization cross linking of recombinant cuticular proteins upon their laccase catalyzed oxidative conjugation with catechols
Insect Biochemistry and Molecular Biology, 2006Co-Authors: Richard J Suderman, Neal T Dittmer, Michael R Kanost, Karl J KramerAbstract:The quinone-tanning hypothesis for insect cuticle Sclerotization proposes that N-acylcatecholamines are oxidized by a phenoloxidase to quinones and quinone methides, which serve as electrophilic cross-linking agents to form covalent cross-links between cuticular proteins. We investigated model reactions for protein cross-linking that occurs during insect cuticle Sclerotization using recombinant pupal cuticular proteins from the tobacco hornworm, Manduca sexta, fungal or recombinant hornworm laccase-type phenoloxidase, and the cross-linking agent precursor N-acylcatecholamines, N-b-alanydopamine (NBAD) or N-acetyldopamine (NADA). Recombinant M. sexta pupal cuticular proteins MsCP36, MsCP20, and MsCP27 were expressed and purified to near homogeneity. Polyclonal antisera to these recombinant proteins recognized the native proteins in crude pharate brown-colored pupal cuticle homogenates. Furthermore, antisera to MsCP36, which contains a type-1 Rebers and Riddiford (RR-1) consensus sequence, also recognized an immunoreactive protein in homogenates of larval head capsule exuviae, indicating the presence of an RR-1 cuticular protein in a very hard, sclerotized and nonpigmented cuticle. All three of the proteins formed small and large oligomers stable to boiling SDS treatment under reducing conditions after reaction with laccase and the N-acylcatecholamines. The optimal reaction conditions for MsCP36 polymerization were 0.3 mM MsCP36, 7.4 mM NBAD and 1.0 U/ml fungal laccase. Approximately 5–10% of the monomer reacted to yield insoluble oligomers and polymers during the reaction, and the monomer also became increasingly insoluble in SDS solution after reaction with the oxidized NBAD. When NADA was used instead of NBAD, less oligomer formation occurred, and most of the protein remained soluble. Radiolabeled NADA became covalently bound to the MsCP36 monomer and oligomers during cross-linking. Recombinant Manduca laccase (MsLac2) also catalyzed the polymerization of MsCP36. These results support the hypothesis that during Sclerotization, insect cuticular proteins are oxidatively conjugated with catechols, a posttranslational process termed catecholation, and then become crosslinked, forming oligomers and subsequently polymers. r 2006 Elsevier Ltd. All rights reserved.
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model reactions for insect cuticle Sclerotization cross linking of recombinant cuticular proteins upon their laccase catalyzed oxidative conjugation with catechols
Insect Biochemistry and Molecular Biology, 2006Co-Authors: Richard J Suderman, Neal T Dittmer, Michael R Kanost, Karl J KramerAbstract:The quinone-tanning hypothesis for insect cuticle Sclerotization proposes that N-acylcatecholamines are oxidized by a phenoloxidase to quinones and quinone methides, which serve as electrophilic cross-linking agents to form covalent cross-links between cuticular proteins. We investigated model reactions for protein cross-linking that occurs during insect cuticle Sclerotization using recombinant pupal cuticular proteins from the tobacco hornworm, Manduca sexta, fungal or recombinant hornworm laccase-type phenoloxidase, and the cross-linking agent precursor N-acylcatecholamines, N-beta-alanydopamine (NBAD) or N-acetyldopamine (NADA). Recombinant M. sexta pupal cuticular proteins MsCP36, MsCP20, and MsCP27 were expressed and purified to near homogeneity. Polyclonal antisera to these recombinant proteins recognized the native proteins in crude pharate brown-colored pupal cuticle homogenates. Furthermore, antisera to MsCP36, which contains a type-1 Rebers and Riddiford (RR-1) consensus sequence, also recognized an immunoreactive protein in homogenates of larval head capsule exuviae, indicating the presence of an RR-1 cuticular protein in a very hard, sclerotized and nonpigmented cuticle. All three of the proteins formed small and large oligomers stable to boiling SDS treatment under reducing conditions after reaction with laccase and the N-acylcatecholamines. The optimal reaction conditions for MsCP36 polymerization were 0.3mM MsCP36, 7.4mM NBAD and 1.0U/mul fungal laccase. Approximately 5-10% of the monomer reacted to yield insoluble oligomers and polymers during the reaction, and the monomer also became increasingly insoluble in SDS solution after reaction with the oxidized NBAD. When NADA was used instead of NBAD, less oligomer formation occurred, and most of the protein remained soluble. Radiolabeled NADA became covalently bound to the MsCP36 monomer and oligomers during cross-linking. Recombinant Manduca laccase (MsLac2) also catalyzed the polymerization of MsCP36. These results support the hypothesis that during Sclerotization, insect cuticular proteins are oxidatively conjugated with catechols, a posttranslational process termed catecholation, and then become cross-linked, forming oligomers and subsequently polymers.
Svend Olav Andersen - One of the best experts on this subject based on the ideXlab platform.
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insect cuticular Sclerotization a review
Insect Biochemistry and Molecular Biology, 2010Co-Authors: Svend Olav AndersenAbstract:Different regions of an insect cuticle have different mechanical properties, partly due to different degrees of stabilization and hardening occurring during the process of Sclerotization, whereby phenolic material is incorporated into the cuticular proteins. Our understanding of the chemistry of cuticular Sclerotization has increased considerably since Mark Pryor in 1940 suggested that enzymatically generated ortho-quinones react with free amino groups, thereby crosslinking the cuticular proteins. The results obtained since then have confirmed the essential features of Pryor's suggestion, and the many observations and experiments, which have been obtained, have led to a detailed and rather complex picture of the Sclerotization process, as described in this review. However, many important questions still remain unanswered, especially regarding the precise regional and temporal regulation of the various steps in the process.
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involvement of tyrosine residues n terminal amino acids and β alanine in insect cuticular Sclerotization
Insect Biochemistry and Molecular Biology, 2007Co-Authors: Svend Olav AndersenAbstract:During Sclerotization of insect cuticle the acyldopamines, N-acetyldopamine (NADA) and N-beta-alanyldopamine (NBAD), are oxidatively incorporated into the cuticular matrix, thereby hardening and stabilizing the material by forming crosslinks between the proteins in the cuticular matrix and by forming polymers filling the intermolecular spaces in the cuticle. Sclerotized cuticle from the locust, Schistocerca gregaria, and the beetle, Tenebrio molitor, was hydrolyzed in dilute hydrochloric acid, and from the hydrolysates some components presumably degradation products of cuticular crosslinks were isolated. In two of the components, the sidechain of 3,4-dihydroxyacetophenone was linked to the amino groups of glycine and beta-alanine, respectively, and in the third component to the phenolic group of tyrosine. These three compounds, glycino-dihydroxyacetophenone, beta-alanino-dihydroxyacetophenone, and O-tyrosino-dihydroxyacetophenone, as well as the previously reported compound, lysino-dihydroxyacetophenone [Andersen, S.O., Roepstorff, P., 2007. Aspects of cuticular Sclerotization in the locust, Schistocerca gregaria, and the beetle, Tenebrio molitor. Insect Biochem. Mol. Biol. 37, 223-234], are suggested to be degradation products of cuticular crosslinks, in which amino acid residues formed linkages to both the alpha- and beta-positions of the sidechain of acyldopamines.
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aspects of cuticular Sclerotization in the locust scistocerca gregaria and the beetle tenebrio molitor
Insect Biochemistry and Molecular Biology, 2007Co-Authors: Svend Olav Andersen, Peter RoepstorffAbstract:Abstract The number of reactive amino groups in cuticular proteins decreases during the early period of insect cuticular Sclerotization, presumably due to reaction with oxidation products of N -acetyldopamine (NADA) and N - β -alanyldopamine (NBAD). We have quantitated the decrease in cuticular N-terminal amino groups and lysine e -amino groups during the first 24 h of Sclerotization in adult locusts, Schistocerca gregaria , and in larval and adult beetles, Tenebrio molitor , as well as the increase in β -alanine amino groups in Tenebrio cuticle. The results indicate that nearly all glycine N-terminal groups and a significant part of the e -amino groups from lysine residues are involved in the Sclerotization process in both locusts and Tenebrio . A pronounced increase in the amount of free β -alanine amino groups was observed in cuticle from adult Tenebrio and to a lesser extent also in Tenebrio larval cuticle, but from locust cuticle no β -alanine was obtained. Hydrolysis of sclerotized cuticles from locusts and Tenebrio by dilute hydrochloric acid released a large number of compounds containing amino acids linked to catecholic moieties. Products have been identified which contain histidine residues linked via their imidazole group to the β -position of various catechols, such as dopamine, 3,4-dihydroxyphenyl-ethanol (DOPET), and 3,4-dihydroxyphenyl-acetaldehyde (DOPALD), and a ketocatecholic compound has also been identified composed of lysine linked via its e -amino group to the α -carbon atom of 3,4-dihydroxyacetophenone. Some of the hydrolysis products have previously been obtained from sclerotized pupal cuticle of Manduca sexta [Xu, R., Huang, X., Hopkins, T.L., Kramer, K.J., 1997. Catecholamine and histidyl protein cross-linked structures in sclerotized insect cuticle. Insect Biochemistry and Molecular Biology 27, 101–108; Kerwin, J.L., Turecek, F., Xu, R., Kramer, K.J., Hopkins, T.L., Gatlin, C.L., Yates, J.R., 1999. Mass spectrometric analysis of catechol-histidine adducts from insect cuticle. Analytical Biochemistry 268, 229–237; Kramer, K.J., Kanost, M.R., Hopkins, T.L., Jiang, H., Zhu, Y.C., Xu, R., Kerwin, J.L., Turecek, F., 2001. Oxidative conjugation of catechols with proteins in insect skeletal systems. Tetrahedron 57, 385–392], but the lysine-dihydroxyacetophenone compound and the histidine–DOPALD adduct have not been reported before. It is suggested that the compounds are derived from NADA and NBAD residues which were incorporated into the cuticle during Sclerotization, and that the lysine-dihydroxyacetophenone as well as the DOPET and DOPALD containing adducts are degradation products derived from cross-links between the cuticular proteins, whereas the dopamine-containing adducts are derived from a non-crosslinking reaction product.
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aspects of cuticular Sclerotization in the locust scistocerca gregaria and the beetle tenebrio molitor
Insect Biochemistry and Molecular Biology, 2007Co-Authors: Svend Olav Andersen, Peter RoepstorffAbstract:The number of reactive amino groups in cuticular proteins decreases during the early period of insect cuticular Sclerotization, presumably due to reaction with oxidation products of N-acetyldopamine (NADA) and N-beta-alanyldopamine (NBAD). We have quantitated the decrease in cuticular N-terminal amino groups and lysine epsilon-amino groups during the first 24h of Sclerotization in adult locusts, Schistocerca gregaria, and in larval and adult beetles, Tenebrio molitor, as well as the increase in beta-alanine amino groups in Tenebrio cuticle. The results indicate that nearly all glycine N-terminal groups and a significant part of the epsilon-amino groups from lysine residues are involved in the Sclerotization process in both locusts and Tenebrio. A pronounced increase in the amount of free beta-alanine amino groups was observed in cuticle from adult Tenebrio and to a lesser extent also in Tenebrio larval cuticle, but from locust cuticle no beta-alanine was obtained. Hydrolysis of sclerotized cuticles from locusts and Tenebrio by dilute hydrochloric acid released a large number of compounds containing amino acids linked to catecholic moieties. Products have been identified which contain histidine residues linked via their imidazole group to the beta-position of various catechols, such as dopamine, 3,4-dihydroxyphenyl-ethanol (DOPET), and 3,4-dihydroxyphenyl-acetaldehyde (DOPALD), and a ketocatecholic compound has also been identified composed of lysine linked via its epsilon-amino group to the alpha-carbon atom of 3,4-dihydroxyacetophenone. Some of the hydrolysis products have previously been obtained from sclerotized pupal cuticle of Manduca sexta [Xu, R., Huang, X., Hopkins, T.L., Kramer, K.J., 1997. Catecholamine and histidyl protein cross-linked structures in sclerotized insect cuticle. Insect Biochemistry and Molecular Biology 27, 101-108; Kerwin, J.L., Turecek, F., Xu, R., Kramer, K.J., Hopkins, T.L., Gatlin, C.L., Yates, J.R., 1999. Mass spectrometric analysis of catechol-histidine adducts from insect cuticle. Analytical Biochemistry 268, 229-237; Kramer, K.J., Kanost, M.R., Hopkins, T.L., Jiang, H., Zhu, Y.C., Xu, R., Kerwin, J.L., Turecek, F., 2001. Oxidative conjugation of catechols with proteins in insect skeletal systems. Tetrahedron 57, 385-392], but the lysine-dihydroxyacetophenone compound and the histidine-DOPALD adduct have not been reported before. It is suggested that the compounds are derived from NADA and NBAD residues which were incorporated into the cuticle during Sclerotization, and that the lysine-dihydroxyacetophenone as well as the DOPET and DOPALD containing adducts are degradation products derived from cross-links between the cuticular proteins, whereas the dopamine-containing adducts are derived from a non-crosslinking reaction product.
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cuticular Sclerotization in insects
Comparative Biochemistry and Physiology B, 1996Co-Authors: Svend Olav Andersen, Martin G Peter, Peter RoepstorffAbstract:Abstract The insect cuticle is an extracellular structure covering the total outer surface of the animal and providing protection against harmful influences from the environment. The mechanical properties of cuticles may vary considerably, and pronounced regional differences are generally observed. The properties may also change during development, and it can be assumed that the physical and chemical properties of all cuticular regions tend to be close to the optimal for proper physiological function during all developmental stages. Cuticular regions can be stabilized by the process of Sclerotization, whereby o-diphenols are oxidatively incorporated into the material. Our current knowledge of the Sclerotization process is reviewed, and it is suggested that the main features of the chemistry of Sclerotization probably have been established, and that the major questions now remaining concern the precise regional and temporal control of the process.
Neal T Dittmer - One of the best experts on this subject based on the ideXlab platform.
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model reactions for insect cuticle Sclerotization participation of amino groups in the cross linking of manduca sexta cuticle protein mscp36
Insect Biochemistry and Molecular Biology, 2010Co-Authors: Richard J Suderman, Neal T Dittmer, Karl J Kramer, Michael R KanostAbstract:Current theories of Sclerotization center on protein cross-linking and dehydration as major factors in the hardening and stability of the insect cuticle. Several studies have reported the identification of catecholamino acid adducts from sclerotizing cuticle involving histidine, lysine, and tyrosine, though there have been no reports of a catechol linked between two amino acid residues. Previously, we reported an in vitro model system for Sclerotization and observed that stable protein oligomers were formed, presumably through cross-links with oxidized catecholamines [Insect Biochem. Mol. Biol. (2006) 36, 353e365]. Using site-directed mutagenesis we created a mutant lacking histidine, rMsCP36(H65A/H178A), to investigate the possible involvement of the two histidine residues of MsCP36 in cross-linking. Surprisingly, this alteration had little ornoeffecton the formation of proteinoligomers as determined bySDS-PAGEanalysis. Blockingof the freeaminogroupsinlysylsidechainsandtheamino-terminus bysuccinylationdiminished, but did not eliminate, cross-linking of either rMsCP36 or rMsCP36(H65A/H178A). We also examined the possibility that cross-linking was due to intermolecular dityrosine linkages. Immunoblot analysis utilizing a monoclonal antibody known to recognize peptidyl dityrosine indicated that dityrosyl cross-links were present. Taken together, these results indicate that lysyl residues are important for the cross-linking of the cuticle protein rMsCP36, but that additional residues other than histidine can also contribute. 2010 Elsevier Ltd. All rights reserved.
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erratumerratum to model reactions for insect cuticle Sclerotization cross linking of recombinant cuticular proteins upon their laccase catalyzed oxidative conjugation with catechols insect biochemistry and molecular biology 36 2006 353 365
Insect Biochemistry and Molecular Biology, 2006Co-Authors: Richard J Suderman, Neal T Dittmer, Michael R Kanost, Karl J KramerAbstract:Erratum to ‘‘Model reactions for insect cuticle Sclerotization: Cross-linking of recombinant cuticular proteins upon their laccase-catalyzed oxidative conjugation with catechols’’ [Insect Biochemistry and Molecular Biology 36 (2006) 353–365] Richard J. Suderman, Neal T. Dittmer, Michael R. Kanost , Karl J. Kramer Department of Biochemistry, 103 Willard Hall, Kansas State University, Manhattan, KS 66506-3706, USA Grain Marketing and Production Research Center, Agricultural Research Service, US Department of Agriculture, Manhattan, KS 66506-2736, USA
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model reactions for insect cuticle Sclerotization cross linking of recombinant cuticular proteins upon their laccase catalyzed oxidative conjugation with catechols
Insect Biochemistry and Molecular Biology, 2006Co-Authors: Richard J Suderman, Neal T Dittmer, Michael R Kanost, Karl J KramerAbstract:The quinone-tanning hypothesis for insect cuticle Sclerotization proposes that N-acylcatecholamines are oxidized by a phenoloxidase to quinones and quinone methides, which serve as electrophilic cross-linking agents to form covalent cross-links between cuticular proteins. We investigated model reactions for protein cross-linking that occurs during insect cuticle Sclerotization using recombinant pupal cuticular proteins from the tobacco hornworm, Manduca sexta, fungal or recombinant hornworm laccase-type phenoloxidase, and the cross-linking agent precursor N-acylcatecholamines, N-b-alanydopamine (NBAD) or N-acetyldopamine (NADA). Recombinant M. sexta pupal cuticular proteins MsCP36, MsCP20, and MsCP27 were expressed and purified to near homogeneity. Polyclonal antisera to these recombinant proteins recognized the native proteins in crude pharate brown-colored pupal cuticle homogenates. Furthermore, antisera to MsCP36, which contains a type-1 Rebers and Riddiford (RR-1) consensus sequence, also recognized an immunoreactive protein in homogenates of larval head capsule exuviae, indicating the presence of an RR-1 cuticular protein in a very hard, sclerotized and nonpigmented cuticle. All three of the proteins formed small and large oligomers stable to boiling SDS treatment under reducing conditions after reaction with laccase and the N-acylcatecholamines. The optimal reaction conditions for MsCP36 polymerization were 0.3 mM MsCP36, 7.4 mM NBAD and 1.0 U/ml fungal laccase. Approximately 5–10% of the monomer reacted to yield insoluble oligomers and polymers during the reaction, and the monomer also became increasingly insoluble in SDS solution after reaction with the oxidized NBAD. When NADA was used instead of NBAD, less oligomer formation occurred, and most of the protein remained soluble. Radiolabeled NADA became covalently bound to the MsCP36 monomer and oligomers during cross-linking. Recombinant Manduca laccase (MsLac2) also catalyzed the polymerization of MsCP36. These results support the hypothesis that during Sclerotization, insect cuticular proteins are oxidatively conjugated with catechols, a posttranslational process termed catecholation, and then become crosslinked, forming oligomers and subsequently polymers. r 2006 Elsevier Ltd. All rights reserved.
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model reactions for insect cuticle Sclerotization cross linking of recombinant cuticular proteins upon their laccase catalyzed oxidative conjugation with catechols
Insect Biochemistry and Molecular Biology, 2006Co-Authors: Richard J Suderman, Neal T Dittmer, Michael R Kanost, Karl J KramerAbstract:The quinone-tanning hypothesis for insect cuticle Sclerotization proposes that N-acylcatecholamines are oxidized by a phenoloxidase to quinones and quinone methides, which serve as electrophilic cross-linking agents to form covalent cross-links between cuticular proteins. We investigated model reactions for protein cross-linking that occurs during insect cuticle Sclerotization using recombinant pupal cuticular proteins from the tobacco hornworm, Manduca sexta, fungal or recombinant hornworm laccase-type phenoloxidase, and the cross-linking agent precursor N-acylcatecholamines, N-b-alanydopamine (NBAD) or N-acetyldopamine (NADA). Recombinant M. sexta pupal cuticular proteins MsCP36, MsCP20, and MsCP27 were expressed and purified to near homogeneity. Polyclonal antisera to these recombinant proteins recognized the native proteins in crude pharate brown-colored pupal cuticle homogenates. Furthermore, antisera to MsCP36, which contains a type-1 Rebers and Riddiford (RR-1) consensus sequence, also recognized an immunoreactive protein in homogenates of larval head capsule exuviae, indicating the presence of an RR-1 cuticular protein in a very hard, sclerotized and nonpigmented cuticle. All three of the proteins formed small and large oligomers stable to boiling SDS treatment under reducing conditions after reaction with laccase and the N-acylcatecholamines. The optimal reaction conditions for MsCP36 polymerization were 0.3 mM MsCP36, 7.4 mM NBAD and 1.0 U/ml fungal laccase. Approximately 5–10% of the monomer reacted to yield insoluble oligomers and polymers during the reaction, and the monomer also became increasingly insoluble in SDS solution after reaction with the oxidized NBAD. When NADA was used instead of NBAD, less oligomer formation occurred, and most of the protein remained soluble. Radiolabeled NADA became covalently bound to the MsCP36 monomer and oligomers during cross-linking. Recombinant Manduca laccase (MsLac2) also catalyzed the polymerization of MsCP36. These results support the hypothesis that during Sclerotization, insect cuticular proteins are oxidatively conjugated with catechols, a posttranslational process termed catecholation, and then become crosslinked, forming oligomers and subsequently polymers. r 2006 Elsevier Ltd. All rights reserved.
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model reactions for insect cuticle Sclerotization cross linking of recombinant cuticular proteins upon their laccase catalyzed oxidative conjugation with catechols
Insect Biochemistry and Molecular Biology, 2006Co-Authors: Richard J Suderman, Neal T Dittmer, Michael R Kanost, Karl J KramerAbstract:The quinone-tanning hypothesis for insect cuticle Sclerotization proposes that N-acylcatecholamines are oxidized by a phenoloxidase to quinones and quinone methides, which serve as electrophilic cross-linking agents to form covalent cross-links between cuticular proteins. We investigated model reactions for protein cross-linking that occurs during insect cuticle Sclerotization using recombinant pupal cuticular proteins from the tobacco hornworm, Manduca sexta, fungal or recombinant hornworm laccase-type phenoloxidase, and the cross-linking agent precursor N-acylcatecholamines, N-beta-alanydopamine (NBAD) or N-acetyldopamine (NADA). Recombinant M. sexta pupal cuticular proteins MsCP36, MsCP20, and MsCP27 were expressed and purified to near homogeneity. Polyclonal antisera to these recombinant proteins recognized the native proteins in crude pharate brown-colored pupal cuticle homogenates. Furthermore, antisera to MsCP36, which contains a type-1 Rebers and Riddiford (RR-1) consensus sequence, also recognized an immunoreactive protein in homogenates of larval head capsule exuviae, indicating the presence of an RR-1 cuticular protein in a very hard, sclerotized and nonpigmented cuticle. All three of the proteins formed small and large oligomers stable to boiling SDS treatment under reducing conditions after reaction with laccase and the N-acylcatecholamines. The optimal reaction conditions for MsCP36 polymerization were 0.3mM MsCP36, 7.4mM NBAD and 1.0U/mul fungal laccase. Approximately 5-10% of the monomer reacted to yield insoluble oligomers and polymers during the reaction, and the monomer also became increasingly insoluble in SDS solution after reaction with the oxidized NBAD. When NADA was used instead of NBAD, less oligomer formation occurred, and most of the protein remained soluble. Radiolabeled NADA became covalently bound to the MsCP36 monomer and oligomers during cross-linking. Recombinant Manduca laccase (MsLac2) also catalyzed the polymerization of MsCP36. These results support the hypothesis that during Sclerotization, insect cuticular proteins are oxidatively conjugated with catechols, a posttranslational process termed catecholation, and then become cross-linked, forming oligomers and subsequently polymers.