The Experts below are selected from a list of 10320 Experts worldwide ranked by ideXlab platform
Kuniyoshi Shimakura - One of the best experts on this subject based on the ideXlab platform.
-
sarcoplasmic calcium binding protein identification as a new allergen of the black tiger shrimp penaeus monodon
International Archives of Allergy and Immunology, 2008Co-Authors: Kazuo Shiomi, Yuichiro Sato, Shohei Hamamoto, Hajime Mita, Kuniyoshi ShimakuraAbstract:Background: Tropomyosin and Arginine Kinase have been identified as crustacean allergens. During purification of Arginine Kinase from black tiger shrimp Penaeus monodon, we found a new allergen of 20-kDa. Methods: A 20-kDa allergen was purified from the abdominal muscle of black tiger shrimp by salting-out, anion-exchange HPLC and reverse-phase HPLC. Following digestion of the 20-kDa allergen with lysyl endopeptidase, peptide fragments were isolated by reverse-phase HPLC, and 2 of them were sequenced. The 20-kDa allergen, together with tropomyosin and Arginine Kinase purified from black tiger shrimp, was evaluated for IgE reactivity by ELISA. Five species of crustaceans (kuruma shrimp, American lobster, pink shrimp, king crab and snow crab) were surveyed for the 20-kDa allergen by immunoblotting. Results: The 20-kDa allergen was purified from black tiger shrimp and identified as a sarcoplasmic calcium-binding protein (SCP) based on the determined amino acid sequences of 2 enzymatic fragments. Of 16 sera from crustacean-allergic patients, 8 and 13 reacted to SCP and tropomyosin, respectively; the reactivity to Arginine Kinase was weakly recognized with 10 sera. In immunoblotting, an IgE-reactive 20-kDa protein was also detected in kuruma shrimp, American lobster and pink shrimp but not in 2 species of crab. Preadsorption of the sera with black tiger shrimp SCP abolished the IgE reactivity of the 20-kDa protein, suggesting the 20-kDa protein to be an SCP. Conclusions: SCP is a new crustacean allergen, and distribution of IgE-reactive SCP is probably limited to shrimp and crayfish.
-
sarcoplasmic calcium binding protein identification as a new allergen of the black tiger shrimp penaeus monodon
International Archives of Allergy and Immunology, 2008Co-Authors: Kazuo Shiomi, Yuichiro Sato, Shohei Hamamoto, Hajime Mita, Kuniyoshi ShimakuraAbstract:Background: Tropomyosin and Arginine Kinase have been identified as crustacean allergens. During purification of Arginine Kinase from black tiger shrimp Penaeus monodon, we found a new allergen of 20-kDa. Methods: A 20-kDa allergen was purified from the abdominal muscle of black tiger shrimp by salting-out, anion-exchange HPLC and reverse-phase HPLC. Following digestion of the 20-kDa allergen with lysyl endopeptidase, peptide fragments were isolated by reverse-phase HPLC, and 2 of them were sequenced. The 20-kDa allergen, together with tropomyosin and Arginine Kinase purified from black tiger shrimp, was evaluated for IgE reactivity by ELISA. Five species of crustaceans (kuruma shrimp, American lobster, pink shrimp, king crab and snow crab) were surveyed for the 20-kDa allergen by immunoblotting. Results: The 20-kDa allergen was purified from black tiger shrimp and identified as a sarcoplasmic calcium-binding protein (SCP) based on the determined amino acid sequences of 2 enzymatic fragments. Of 16 sera from crustacean-allergic patients, 8 and 13 reacted to SCP and tropomyosin, respectively; the reactivity to Arginine Kinase was weakly recognized with 10 sera. In immunoblotting, an IgE-reactive 20-kDa protein was also detected in kuruma shrimp, American lobster and pink shrimp but not in 2 species of crab. Preadsorption of the sera with black tiger shrimp SCP abolished the IgE reactivity of the 20-kDa protein, suggesting the 20-kDa protein to be an SCP. Conclusions: SCP is a new crustacean allergen, and distribution of IgE-reactive SCP is probably limited to shrimp and crayfish.
Kazuo Shiomi - One of the best experts on this subject based on the ideXlab platform.
-
sarcoplasmic calcium binding protein identification as a new allergen of the black tiger shrimp penaeus monodon
International Archives of Allergy and Immunology, 2008Co-Authors: Kazuo Shiomi, Yuichiro Sato, Shohei Hamamoto, Hajime Mita, Kuniyoshi ShimakuraAbstract:Background: Tropomyosin and Arginine Kinase have been identified as crustacean allergens. During purification of Arginine Kinase from black tiger shrimp Penaeus monodon, we found a new allergen of 20-kDa. Methods: A 20-kDa allergen was purified from the abdominal muscle of black tiger shrimp by salting-out, anion-exchange HPLC and reverse-phase HPLC. Following digestion of the 20-kDa allergen with lysyl endopeptidase, peptide fragments were isolated by reverse-phase HPLC, and 2 of them were sequenced. The 20-kDa allergen, together with tropomyosin and Arginine Kinase purified from black tiger shrimp, was evaluated for IgE reactivity by ELISA. Five species of crustaceans (kuruma shrimp, American lobster, pink shrimp, king crab and snow crab) were surveyed for the 20-kDa allergen by immunoblotting. Results: The 20-kDa allergen was purified from black tiger shrimp and identified as a sarcoplasmic calcium-binding protein (SCP) based on the determined amino acid sequences of 2 enzymatic fragments. Of 16 sera from crustacean-allergic patients, 8 and 13 reacted to SCP and tropomyosin, respectively; the reactivity to Arginine Kinase was weakly recognized with 10 sera. In immunoblotting, an IgE-reactive 20-kDa protein was also detected in kuruma shrimp, American lobster and pink shrimp but not in 2 species of crab. Preadsorption of the sera with black tiger shrimp SCP abolished the IgE reactivity of the 20-kDa protein, suggesting the 20-kDa protein to be an SCP. Conclusions: SCP is a new crustacean allergen, and distribution of IgE-reactive SCP is probably limited to shrimp and crayfish.
-
sarcoplasmic calcium binding protein identification as a new allergen of the black tiger shrimp penaeus monodon
International Archives of Allergy and Immunology, 2008Co-Authors: Kazuo Shiomi, Yuichiro Sato, Shohei Hamamoto, Hajime Mita, Kuniyoshi ShimakuraAbstract:Background: Tropomyosin and Arginine Kinase have been identified as crustacean allergens. During purification of Arginine Kinase from black tiger shrimp Penaeus monodon, we found a new allergen of 20-kDa. Methods: A 20-kDa allergen was purified from the abdominal muscle of black tiger shrimp by salting-out, anion-exchange HPLC and reverse-phase HPLC. Following digestion of the 20-kDa allergen with lysyl endopeptidase, peptide fragments were isolated by reverse-phase HPLC, and 2 of them were sequenced. The 20-kDa allergen, together with tropomyosin and Arginine Kinase purified from black tiger shrimp, was evaluated for IgE reactivity by ELISA. Five species of crustaceans (kuruma shrimp, American lobster, pink shrimp, king crab and snow crab) were surveyed for the 20-kDa allergen by immunoblotting. Results: The 20-kDa allergen was purified from black tiger shrimp and identified as a sarcoplasmic calcium-binding protein (SCP) based on the determined amino acid sequences of 2 enzymatic fragments. Of 16 sera from crustacean-allergic patients, 8 and 13 reacted to SCP and tropomyosin, respectively; the reactivity to Arginine Kinase was weakly recognized with 10 sera. In immunoblotting, an IgE-reactive 20-kDa protein was also detected in kuruma shrimp, American lobster and pink shrimp but not in 2 species of crab. Preadsorption of the sera with black tiger shrimp SCP abolished the IgE reactivity of the 20-kDa protein, suggesting the 20-kDa protein to be an SCP. Conclusions: SCP is a new crustacean allergen, and distribution of IgE-reactive SCP is probably limited to shrimp and crayfish.
Tomohiko Suzuki - One of the best experts on this subject based on the ideXlab platform.
-
evidence for n terminal myristoylation of tetrahymena Arginine Kinase using peptide mass fingerprinting analysis
Protein Journal, 2016Co-Authors: Shou Motomura, Tomohiko SuzukiAbstract:In this study, we confirmed N-terminal myristoylation of Tetrahymena pyriformis Arginine Kinase (AK1) by identifying a myristoylation signal sequence at the N-terminus. A sufficient amount of modified enzyme was synthesized using an insect cell-free protein synthesis system that contains all of the elements necessary for post-transcriptional modification by fatty acids. Subsequent peptide mass fingerprinting (PMF) analyses were performed after digestion with trypsin. The PMF data covered 39 % (143 residues) of internal peptides. The target N-myristoylated peptide had a theoretical mass of 832.4477 and was clearly observed with an experimental mass (m/z-H(+)) of 832.4747. The difference between the two masses was 0.0271, supporting the accuracy of identification and indicating that the synthesized T. pyriformis AK1 is myristoylated. The fixed specimens of T. pyriformis were reacted with an anti-AK1 peptide antibody followed by a secondary antibody with a fluorescent chromophore and were observed using immunofluorescence microscope. In agreement with previous western blotting analyses, microscopic observations suggested that AK1 is localized in the cilia. The present PMF and microscopic analyses indicate that T. pyriformis AK1 may be localized and anchored to ciliary membranes via N-terminal myristoyl groups.
-
characterization of Arginine Kinase from the caenogastropod semisulcospira libertina an intermediate host of paragonimus westermani
Journal of Molluscan Studies, 2014Co-Authors: Blanca R Jarilla, Luz P Acosta, Tomohiko Suzuki, Misako Urabe, Tomonori AgatsumaAbstract:Arginine Kinase (AK) belongs to the phosphagen Kinase (PK) family of enzymes that have a significant role in the maintenance of energy homeostasis. In this study, cDNA of Semisulcospira libertina AK was determined, cloned and the recombinant protein was expressed as fusion protein with maltose-binding protein. The protein has a theoretical molecular mass of 39,412 Da and an estimated isoelectric point (p I) of 6.37. The recombinant enzyme showed high affinity and significant activity only for the substrate L-Arginine (Km ¼ 0.53 mM; Vmax ¼ 61.30 mmol/min mg protein). Phylogenetic analyses showed that S. libertina AK clusters with gastropod and cephalopod AKs. Comparison of gene structure showed that all intron positions of S. libertina AK are shared with molluscan AKs. Certain intron positions are also shared with trematode TK and sipunculid HTK. Semisulcospira libertina AK can be utilized in the control of paragonimiasis since it is absent in mammalian hosts.
-
gene structure of the two domain taurocyamine Kinase from paragonimus westermani evidence for a distinct lineage of trematode phosphagen Kinases
FEBS Letters, 2013Co-Authors: Mitsuru Nagataki, Blanca R Jarilla, Shinji Tokuhiro, Luz P Acosta, Tomohiko Suzuki, Tomonori AgatsumaAbstract:Taurocyamine Kinase (TK) is an enzyme that catalyzes the reversible transfer of a phosphate between ATP and taurocyamine. Annelid TKs were suggested to have evolved from a CK ancestor. However, TKs from the lung fluke Paragonimus westermani comprised another lineage. Construction of phylogenetic tree and comparison of exon/intron organization showed that P. westermani TK and other trematode TKs evolved from a molluscan Arginine Kinase (AK) gene. Exon shuffling probably caused the changes in amino acid sequence thereby changing the affinity from AK to TK. The present study provides new insights on the evolution of phosphagen Kinases found in trematodes.
-
development of a highly sensitive igg elisa based on recombinant Arginine Kinase of toxocara canis for serodiagnosis of visceral larva migrans in the murine model
Parasitology Research, 2008Co-Authors: Susiji Wickramasinghe, Mitsuru Nagataki, Tomohiko Suzuki, Lalani Yatawara, R P V J Rajapakse, Yoshiya Watanabe, Misa Takamoto, Tomonori AgatsumaAbstract:Toxocariasis is a worldwide zoonotic disease caused by the ascarid nematode Toxocara canis. The most common method available for serodiagnosis of toxocariasis is an enzyme-linked immunosorbent assay (ELISA) test using Toxocara excretory-secretory antigen (TES). The present study describes the development of IgG-ELISA based on antiserum prepared against the recombinant Arginine Kinase of Toxocara canis. Antiserum was prepared against the purified recombinant Arginine Kinase (AK) using 6-week-old female Japanese white rabbits. Serum samples were collected from experimentally infected BALB/c and C57BL/6 mice at different time periods. The IgG-ELISA was performed using serum samples from mice (infected/uninfected) and TES antigen with antiserum prepared against the recombinant-AK. The optical density (OD450) was measured at 450 nm using a micro-plate ELISA reader. There were significant differences (P < 0.01) in the absorbance between infected and control serum samples. Further, we obtained 100% sensitivity for the serum samples from T. canis-infected mice. Therefore, it is suggested that the recombinant-AK based IgG-ELISA could be applied for immunodiagnosis of human toxocariasis. However, it is necessary to evaluate the specificity of this recombinant antigen with similar geohelminth infections.
-
toxocara canis molecular cloning characterization expression and comparison of the kinetics of cdna derived Arginine Kinase
Experimental Parasitology, 2007Co-Authors: Susiji Wickramasinghe, Mitsuru Nagataki, Tomohiko Suzuki, Kouji Uda, Lalani Yatawara, R P V J Rajapakse, Yoshiya Watanabe, Tomonori AgatsumaAbstract:Abstract Arginine Kinase (AK) is a member of a highly conserved family of phosphagen Kinases. We determined the cDNA sequence of Toxocara canis AK, cloned it in pMAL plasmid and expressed it in Escherichia coli as a fusion protein with maltose-binding protein. The protein has a theoretical molecular mass of 45,376 Da and an estimated isoelectric point (pI) of 8.38. Alignment of the cDNA-derived amino acid sequence of T. canis AK with other phosphagen Kinase sequences showed high amino acid identity with other nematode AKs, and phylogenetic analysis placed it as a distinct branch within a nematode AK cluster. Analysis of the N-terminus sequence of T. canis AK revealed the presence of a signal targeting peptide presumably targeting this protein to cytosol or endoplasmic reticulum (ER). T. canis AK showed high activity for l -Arginine. The kinetic constants (Km = 0.12 mM, Kcat = 29.18, and Kd = 0.23 mM) and Vmax (43.76 μmol Pi/min/mg protein) of T. canis recombinant-AK were determined for the forward reaction. It also exhibited a synergism for substrate binding ( K d Arg / K m Arg = 1.96 ) . Comparison of K cat / K m Arg values in various Arginine Kinases indicates that T. canis AK has a high catalytic efficiency (248.19 s−1 mM−1). The present study contains the first description of Arginine Kinase in a zoonotic nematode. The determination of T. canis AK and its phosphagen biosynthetic pathway, which is completely different from those in mammalian host tissues, suggests this enzyme as a possible novel chemotherapy target for VLM syndrome in humans.
Hajime Mita - One of the best experts on this subject based on the ideXlab platform.
-
sarcoplasmic calcium binding protein identification as a new allergen of the black tiger shrimp penaeus monodon
International Archives of Allergy and Immunology, 2008Co-Authors: Kazuo Shiomi, Yuichiro Sato, Shohei Hamamoto, Hajime Mita, Kuniyoshi ShimakuraAbstract:Background: Tropomyosin and Arginine Kinase have been identified as crustacean allergens. During purification of Arginine Kinase from black tiger shrimp Penaeus monodon, we found a new allergen of 20-kDa. Methods: A 20-kDa allergen was purified from the abdominal muscle of black tiger shrimp by salting-out, anion-exchange HPLC and reverse-phase HPLC. Following digestion of the 20-kDa allergen with lysyl endopeptidase, peptide fragments were isolated by reverse-phase HPLC, and 2 of them were sequenced. The 20-kDa allergen, together with tropomyosin and Arginine Kinase purified from black tiger shrimp, was evaluated for IgE reactivity by ELISA. Five species of crustaceans (kuruma shrimp, American lobster, pink shrimp, king crab and snow crab) were surveyed for the 20-kDa allergen by immunoblotting. Results: The 20-kDa allergen was purified from black tiger shrimp and identified as a sarcoplasmic calcium-binding protein (SCP) based on the determined amino acid sequences of 2 enzymatic fragments. Of 16 sera from crustacean-allergic patients, 8 and 13 reacted to SCP and tropomyosin, respectively; the reactivity to Arginine Kinase was weakly recognized with 10 sera. In immunoblotting, an IgE-reactive 20-kDa protein was also detected in kuruma shrimp, American lobster and pink shrimp but not in 2 species of crab. Preadsorption of the sera with black tiger shrimp SCP abolished the IgE reactivity of the 20-kDa protein, suggesting the 20-kDa protein to be an SCP. Conclusions: SCP is a new crustacean allergen, and distribution of IgE-reactive SCP is probably limited to shrimp and crayfish.
-
sarcoplasmic calcium binding protein identification as a new allergen of the black tiger shrimp penaeus monodon
International Archives of Allergy and Immunology, 2008Co-Authors: Kazuo Shiomi, Yuichiro Sato, Shohei Hamamoto, Hajime Mita, Kuniyoshi ShimakuraAbstract:Background: Tropomyosin and Arginine Kinase have been identified as crustacean allergens. During purification of Arginine Kinase from black tiger shrimp Penaeus monodon, we found a new allergen of 20-kDa. Methods: A 20-kDa allergen was purified from the abdominal muscle of black tiger shrimp by salting-out, anion-exchange HPLC and reverse-phase HPLC. Following digestion of the 20-kDa allergen with lysyl endopeptidase, peptide fragments were isolated by reverse-phase HPLC, and 2 of them were sequenced. The 20-kDa allergen, together with tropomyosin and Arginine Kinase purified from black tiger shrimp, was evaluated for IgE reactivity by ELISA. Five species of crustaceans (kuruma shrimp, American lobster, pink shrimp, king crab and snow crab) were surveyed for the 20-kDa allergen by immunoblotting. Results: The 20-kDa allergen was purified from black tiger shrimp and identified as a sarcoplasmic calcium-binding protein (SCP) based on the determined amino acid sequences of 2 enzymatic fragments. Of 16 sera from crustacean-allergic patients, 8 and 13 reacted to SCP and tropomyosin, respectively; the reactivity to Arginine Kinase was weakly recognized with 10 sera. In immunoblotting, an IgE-reactive 20-kDa protein was also detected in kuruma shrimp, American lobster and pink shrimp but not in 2 species of crab. Preadsorption of the sera with black tiger shrimp SCP abolished the IgE reactivity of the 20-kDa protein, suggesting the 20-kDa protein to be an SCP. Conclusions: SCP is a new crustacean allergen, and distribution of IgE-reactive SCP is probably limited to shrimp and crayfish.
Shohei Hamamoto - One of the best experts on this subject based on the ideXlab platform.
-
sarcoplasmic calcium binding protein identification as a new allergen of the black tiger shrimp penaeus monodon
International Archives of Allergy and Immunology, 2008Co-Authors: Kazuo Shiomi, Yuichiro Sato, Shohei Hamamoto, Hajime Mita, Kuniyoshi ShimakuraAbstract:Background: Tropomyosin and Arginine Kinase have been identified as crustacean allergens. During purification of Arginine Kinase from black tiger shrimp Penaeus monodon, we found a new allergen of 20-kDa. Methods: A 20-kDa allergen was purified from the abdominal muscle of black tiger shrimp by salting-out, anion-exchange HPLC and reverse-phase HPLC. Following digestion of the 20-kDa allergen with lysyl endopeptidase, peptide fragments were isolated by reverse-phase HPLC, and 2 of them were sequenced. The 20-kDa allergen, together with tropomyosin and Arginine Kinase purified from black tiger shrimp, was evaluated for IgE reactivity by ELISA. Five species of crustaceans (kuruma shrimp, American lobster, pink shrimp, king crab and snow crab) were surveyed for the 20-kDa allergen by immunoblotting. Results: The 20-kDa allergen was purified from black tiger shrimp and identified as a sarcoplasmic calcium-binding protein (SCP) based on the determined amino acid sequences of 2 enzymatic fragments. Of 16 sera from crustacean-allergic patients, 8 and 13 reacted to SCP and tropomyosin, respectively; the reactivity to Arginine Kinase was weakly recognized with 10 sera. In immunoblotting, an IgE-reactive 20-kDa protein was also detected in kuruma shrimp, American lobster and pink shrimp but not in 2 species of crab. Preadsorption of the sera with black tiger shrimp SCP abolished the IgE reactivity of the 20-kDa protein, suggesting the 20-kDa protein to be an SCP. Conclusions: SCP is a new crustacean allergen, and distribution of IgE-reactive SCP is probably limited to shrimp and crayfish.
-
sarcoplasmic calcium binding protein identification as a new allergen of the black tiger shrimp penaeus monodon
International Archives of Allergy and Immunology, 2008Co-Authors: Kazuo Shiomi, Yuichiro Sato, Shohei Hamamoto, Hajime Mita, Kuniyoshi ShimakuraAbstract:Background: Tropomyosin and Arginine Kinase have been identified as crustacean allergens. During purification of Arginine Kinase from black tiger shrimp Penaeus monodon, we found a new allergen of 20-kDa. Methods: A 20-kDa allergen was purified from the abdominal muscle of black tiger shrimp by salting-out, anion-exchange HPLC and reverse-phase HPLC. Following digestion of the 20-kDa allergen with lysyl endopeptidase, peptide fragments were isolated by reverse-phase HPLC, and 2 of them were sequenced. The 20-kDa allergen, together with tropomyosin and Arginine Kinase purified from black tiger shrimp, was evaluated for IgE reactivity by ELISA. Five species of crustaceans (kuruma shrimp, American lobster, pink shrimp, king crab and snow crab) were surveyed for the 20-kDa allergen by immunoblotting. Results: The 20-kDa allergen was purified from black tiger shrimp and identified as a sarcoplasmic calcium-binding protein (SCP) based on the determined amino acid sequences of 2 enzymatic fragments. Of 16 sera from crustacean-allergic patients, 8 and 13 reacted to SCP and tropomyosin, respectively; the reactivity to Arginine Kinase was weakly recognized with 10 sera. In immunoblotting, an IgE-reactive 20-kDa protein was also detected in kuruma shrimp, American lobster and pink shrimp but not in 2 species of crab. Preadsorption of the sera with black tiger shrimp SCP abolished the IgE reactivity of the 20-kDa protein, suggesting the 20-kDa protein to be an SCP. Conclusions: SCP is a new crustacean allergen, and distribution of IgE-reactive SCP is probably limited to shrimp and crayfish.