The Experts below are selected from a list of 231 Experts worldwide ranked by ideXlab platform

Jiang Xia - One of the best experts on this subject based on the ideXlab platform.

  • Versatile Site-Selective Protein Reaction Guided by WW Domain-Peptide Motif Interaction.
    Bioconjugate chemistry, 2017
    Co-Authors: Miao Liu, Mingjie Zhang, Jiang Xia
    Abstract:

    A short, flexible, and unstructured peptide tag that has versatile and facile use in protein labeling applications is highly desirable. Here, we report an 11-residue peptide tag with an internal cysteine (a W-tag, derived from a Comm PY peptide motif that is known to bind with Nedd4 WW3* domain) that can be installed at different regions of the target protein without compromising its covalent reactivity with the reactive label (a 35-residue synthetic Nedd4 WW3* domain derivative). This versatility is explained by the unique structural features of the reaction. NMR analysis reveals that both the W-tag peptide and reactive Nedd4 WW3* protein are unstructured before they encounter each other. The binding interaction of the two induces noticeable structural changes and promotes global folding. Consequently, the reactive cysteine residue at W-tag and the electrophilic Chloroacetyl Group at Nedd4 WW3* domain are positioned to be in close proximity, inducing an intermolecular covalent cross-linking. The covalent...

  • Versatile Site-Selective Protein Reaction Guided by WW Domain–Peptide Motif Interaction
    2017
    Co-Authors: Miao Liu, Mingjie Zhang, Jiang Xia
    Abstract:

    A short, flexible, and unstructured peptide tag that has versatile and facile use in protein labeling applications is highly desirable. Here, we report an 11-residue peptide tag with an internal cysteine (a W-tag, derived from a Comm PY peptide motif that is known to bind with Nedd4 WW3* domain) that can be installed at different regions of the target protein without compromising its covalent reactivity with the reactive label (a 35-residue synthetic Nedd4 WW3* domain derivative). This versatility is explained by the unique structural features of the reaction. NMR analysis reveals that both the W-tag peptide and reactive Nedd4 WW3* protein are unstructured before they encounter each other. The binding interaction of the two induces noticeable structural changes and promotes global folding. Consequently, the reactive cysteine residue at W-tag and the electrophilic Chloroacetyl Group at Nedd4 WW3* domain are positioned to be in close proximity, inducing an intermolecular covalent cross-linking. The covalent linkage in turn stabilizes the folding of the protein complex. This unique multistep mechanism renders this labeling reaction amenable to different sites of the proteins of interest: installation of the tag at N- and C-termini, in the flexible linker region, in the loop region, and the extracellular terminus of target proteins exhibited comparable reactivity. This work therefore represents the first proximity-induced cysteine reaction based on the unique binding features of WW domains that demonstrates unprecedented versatility

  • Affinity-guided covalent conjugation reactions based on PDZ-peptide and SH3-peptide interactions.
    Bioconjugate chemistry, 2014
    Co-Authors: Feng Huang, Jianpeng Wang, Jiang Xia
    Abstract:

    Specific protein-peptide interactions are prevalent in the living cells and form a tightly regulated signaling network. These interactions, many of which have structural information revealed, provide ideal templates for affinity-guided covalent bioconjugation. Here we report the development of a set of four new reactions that covalently and site-specifically link nonenzymatic scaffolding domains (two PDZ and two SH3 domains) and their ligands through thiol-Chloroacetyl SN2 reaction. Guided by the three-dimensional structure of the wild type complex, a selected position of the protein was mutated to cysteine, and at the same time, an α-Chloroacetyl Group was installed at a corresponding position of the peptide. Specific binding interaction between the two brings the reactive Groups into close proximity, converts the nonreactive cysteine residue into a content-dependent reactive site, and induces the nucleophilic reaction that is inert in the absence of the binding event. The specificity, orthogonality, and modularity of the four reactions were characterized, the reaction was applied to label proteins in vitro and receptor on the surface of mammalian cells, and the system was utilized to assemble covalent protein complexes with unnatural geometries.

  • Short Peptide Tag for Covalent Protein Labeling Based on Coiled Coils
    2014
    Co-Authors: Jianpeng Wang, Jiang Xia
    Abstract:

    To label proteins covalently, one faces a trade-off between labeling a protein specifically and using a small tag. Often one must compromise one parameter for the other or use additional components, such as an enzyme, to satisfy both requirements. Here, we report a new reaction that covalently labels proteins by using engineered coiled-coil peptides. Harnessing the concept of “proximity-induced reactivity”, the 21-amino-acid three-heptad peptides CCE/CCK were modified with a nucleophilic cysteine and an α-Chloroacetyl Group at selected positions. When pairs of coiled coils associated, an irreversible covalent bond spontaneously formed between the peptides. The specificity of the cross-linking reaction was characterized, the probes were improved by making them bivalent, and the system was used to label a protein in vitro and receptors on the surface of mammalian cells

  • Affinity-Guided Covalent Conjugation Reactions Based on PDZ–Peptide and SH3–Peptide Interactions
    2014
    Co-Authors: Feng Huang, Jianpeng Wang, Jiang Xia
    Abstract:

    Specific protein–peptide interactions are prevalent in the living cells and form a tightly regulated signaling network. These interactions, many of which have structural information revealed, provide ideal templates for affinity-guided covalent bioconjugation. Here we report the development of a set of four new reactions that covalently and site-specifically link nonenzymatic scaffolding domains (two PDZ and two SH3 domains) and their ligands through thiol-Chloroacetyl SN2 reaction. Guided by the three-dimensional structure of the wild type complex, a selected position of the protein was mutated to cysteine, and at the same time, an α-Chloroacetyl Group was installed at a corresponding position of the peptide. Specific binding interaction between the two brings the reactive Groups into close proximity, converts the nonreactive cysteine residue into a content-dependent reactive site, and induces the nucleophilic reaction that is inert in the absence of the binding event. The specificity, orthogonality, and modularity of the four reactions were characterized, the reaction was applied to label proteins in vitro and receptor on the surface of mammalian cells, and the system was utilized to assemble covalent protein complexes with unnatural geometries

Hiroaki Suga - One of the best experts on this subject based on the ideXlab platform.

  • Synthesis of fused tricyclic peptides using a reprogrammed translation system and chemical modification.
    Bioorganic chemistry, 2015
    Co-Authors: Nasir K. Bashiruddin, Masanobu Nagano, Hiroaki Suga
    Abstract:

    Here we report a unique method of ribosomally synthesizing fused tricyclic peptides. Flexizyme-assisted in vitro translation of a linear peptide with the N-terminal Chloroacetyl Group and four downstream cysteines followed by the addition of 1,3,5-tris(bromomethyl)benzene results in selective production of the fused tricyclic peptide. This technology can be used for the ribosomal synthesis of fused tricyclic peptide libraries for the in vitro selection of bioactive peptides with tricyclic topology.

  • Selective thioether macrocyclization of peptides having the N-terminal 2-Chloroacetyl Group and competing two or three cysteine residues in translation
    Organic & biomolecular chemistry, 2012
    Co-Authors: Kazuhiro Iwasaki, Yuki Goto, Takayuki Katoh, Hiroaki Suga
    Abstract:

    The mode of thioether macrocyclization of peptides containing an N-terminal 2-Chloroacetyl Group and two or three competing cysteine residues at downstream positions has been extensively studied, leading to a strategy for designated formation of overlapping-bicyclic peptides or dumbbell-type bicyclic peptides.

  • Ribosomal Synthesis of Bicyclic Peptides via Two Orthogonal Inter-Side-Chain Reactions
    Journal of the American Chemical Society, 2008
    Co-Authors: Yusuke Sako, Jumpei Morimoto, Hiroshi Murakami, Hiroaki Suga
    Abstract:

    Here we report a new methodology for the synthesis of bicyclic peptides by using a reconstituted cell-free translation system under the reprogrammed genetic code. Cysteine (Cys) and three different nonproteinogenic amino acids, Cab, Aha, and Pgl, were simultaneously incorporated into a peptide chain. The first cyclization occurred between the Chloroacetyl Group of Cab and the sulfhydryl Group in Cys in situ of translation, and the second cyclization on the side chains of Aha−Pgl via Cu(I)-catalyzed azide−alkyne cycloaddition was performed. This offers us a powerful means of mRNA-programmed synthesis of various peptides with uniform bicyclic scaffolds.

  • Ribosomal synthesis of peptidase-resistant peptides closed by a nonreducible inter-side-chain bond.
    ACS chemical biology, 2008
    Co-Authors: Yusuke Sako, Yuki Goto, Hiroshi Murakami, Hiroaki Suga
    Abstract:

    Here we report a new enabling technology for the synthesis of peptidase-resistant cyclic peptides by means of genetic code reprogramming involving the flexizyme (a tRNA acylation ribozyme) and PURE (a reconstituted cell-free translation) systems. In this work, we have developed a new nonproteinogenic amino acid bearing a Chloroacetyl Group in the side chain, which forms a physiologically stable thioether bond by intramolecular reaction with the sulfhydryl Group of a Cys residue in the peptide chain upon translation. Significantly, this chemistry takes place spontaneously in situ of the translation solution, giving the corresponding cyclic peptides independent of ring sizes. We have used this method to convert human urotensin II, known as a potent vasoconstrictor, to its analogue containing a thioether bond, showing that this new analogue retains biological activity. Moreover, this peptide exhibits remarkable resistance against peptidases under reducing conditions. Thus, this technology offers a new means ...

Miao Liu - One of the best experts on this subject based on the ideXlab platform.

  • Versatile Site-Selective Protein Reaction Guided by WW Domain-Peptide Motif Interaction.
    Bioconjugate chemistry, 2017
    Co-Authors: Miao Liu, Mingjie Zhang, Jiang Xia
    Abstract:

    A short, flexible, and unstructured peptide tag that has versatile and facile use in protein labeling applications is highly desirable. Here, we report an 11-residue peptide tag with an internal cysteine (a W-tag, derived from a Comm PY peptide motif that is known to bind with Nedd4 WW3* domain) that can be installed at different regions of the target protein without compromising its covalent reactivity with the reactive label (a 35-residue synthetic Nedd4 WW3* domain derivative). This versatility is explained by the unique structural features of the reaction. NMR analysis reveals that both the W-tag peptide and reactive Nedd4 WW3* protein are unstructured before they encounter each other. The binding interaction of the two induces noticeable structural changes and promotes global folding. Consequently, the reactive cysteine residue at W-tag and the electrophilic Chloroacetyl Group at Nedd4 WW3* domain are positioned to be in close proximity, inducing an intermolecular covalent cross-linking. The covalent...

  • Versatile Site-Selective Protein Reaction Guided by WW Domain–Peptide Motif Interaction
    2017
    Co-Authors: Miao Liu, Mingjie Zhang, Jiang Xia
    Abstract:

    A short, flexible, and unstructured peptide tag that has versatile and facile use in protein labeling applications is highly desirable. Here, we report an 11-residue peptide tag with an internal cysteine (a W-tag, derived from a Comm PY peptide motif that is known to bind with Nedd4 WW3* domain) that can be installed at different regions of the target protein without compromising its covalent reactivity with the reactive label (a 35-residue synthetic Nedd4 WW3* domain derivative). This versatility is explained by the unique structural features of the reaction. NMR analysis reveals that both the W-tag peptide and reactive Nedd4 WW3* protein are unstructured before they encounter each other. The binding interaction of the two induces noticeable structural changes and promotes global folding. Consequently, the reactive cysteine residue at W-tag and the electrophilic Chloroacetyl Group at Nedd4 WW3* domain are positioned to be in close proximity, inducing an intermolecular covalent cross-linking. The covalent linkage in turn stabilizes the folding of the protein complex. This unique multistep mechanism renders this labeling reaction amenable to different sites of the proteins of interest: installation of the tag at N- and C-termini, in the flexible linker region, in the loop region, and the extracellular terminus of target proteins exhibited comparable reactivity. This work therefore represents the first proximity-induced cysteine reaction based on the unique binding features of WW domains that demonstrates unprecedented versatility

Jianpeng Wang - One of the best experts on this subject based on the ideXlab platform.

  • Affinity-guided covalent conjugation reactions based on PDZ-peptide and SH3-peptide interactions.
    Bioconjugate chemistry, 2014
    Co-Authors: Feng Huang, Jianpeng Wang, Jiang Xia
    Abstract:

    Specific protein-peptide interactions are prevalent in the living cells and form a tightly regulated signaling network. These interactions, many of which have structural information revealed, provide ideal templates for affinity-guided covalent bioconjugation. Here we report the development of a set of four new reactions that covalently and site-specifically link nonenzymatic scaffolding domains (two PDZ and two SH3 domains) and their ligands through thiol-Chloroacetyl SN2 reaction. Guided by the three-dimensional structure of the wild type complex, a selected position of the protein was mutated to cysteine, and at the same time, an α-Chloroacetyl Group was installed at a corresponding position of the peptide. Specific binding interaction between the two brings the reactive Groups into close proximity, converts the nonreactive cysteine residue into a content-dependent reactive site, and induces the nucleophilic reaction that is inert in the absence of the binding event. The specificity, orthogonality, and modularity of the four reactions were characterized, the reaction was applied to label proteins in vitro and receptor on the surface of mammalian cells, and the system was utilized to assemble covalent protein complexes with unnatural geometries.

  • Short Peptide Tag for Covalent Protein Labeling Based on Coiled Coils
    2014
    Co-Authors: Jianpeng Wang, Jiang Xia
    Abstract:

    To label proteins covalently, one faces a trade-off between labeling a protein specifically and using a small tag. Often one must compromise one parameter for the other or use additional components, such as an enzyme, to satisfy both requirements. Here, we report a new reaction that covalently labels proteins by using engineered coiled-coil peptides. Harnessing the concept of “proximity-induced reactivity”, the 21-amino-acid three-heptad peptides CCE/CCK were modified with a nucleophilic cysteine and an α-Chloroacetyl Group at selected positions. When pairs of coiled coils associated, an irreversible covalent bond spontaneously formed between the peptides. The specificity of the cross-linking reaction was characterized, the probes were improved by making them bivalent, and the system was used to label a protein in vitro and receptors on the surface of mammalian cells

  • Affinity-Guided Covalent Conjugation Reactions Based on PDZ–Peptide and SH3–Peptide Interactions
    2014
    Co-Authors: Feng Huang, Jianpeng Wang, Jiang Xia
    Abstract:

    Specific protein–peptide interactions are prevalent in the living cells and form a tightly regulated signaling network. These interactions, many of which have structural information revealed, provide ideal templates for affinity-guided covalent bioconjugation. Here we report the development of a set of four new reactions that covalently and site-specifically link nonenzymatic scaffolding domains (two PDZ and two SH3 domains) and their ligands through thiol-Chloroacetyl SN2 reaction. Guided by the three-dimensional structure of the wild type complex, a selected position of the protein was mutated to cysteine, and at the same time, an α-Chloroacetyl Group was installed at a corresponding position of the peptide. Specific binding interaction between the two brings the reactive Groups into close proximity, converts the nonreactive cysteine residue into a content-dependent reactive site, and induces the nucleophilic reaction that is inert in the absence of the binding event. The specificity, orthogonality, and modularity of the four reactions were characterized, the reaction was applied to label proteins in vitro and receptor on the surface of mammalian cells, and the system was utilized to assemble covalent protein complexes with unnatural geometries

  • Short Peptide Tag for Covalent Protein Labeling Based on Coiled Coils
    Bioconjugate chemistry, 2013
    Co-Authors: Jianpeng Wang, Jiang Xia
    Abstract:

    To label proteins covalently, one faces a trade-off between labeling a protein specifically and using a small tag. Often one must compromise one parameter for the other or use additional components, such as an enzyme, to satisfy both requirements. Here, we report a new reaction that covalently labels proteins by using engineered coiled-coil peptides. Harnessing the concept of “proximity-induced reactivity”, the 21-amino-acid three-heptad peptides CCE/CCK were modified with a nucleophilic cysteine and an α-Chloroacetyl Group at selected positions. When pairs of coiled coils associated, an irreversible covalent bond spontaneously formed between the peptides. The specificity of the cross-linking reaction was characterized, the probes were improved by making them bivalent, and the system was used to label a protein in vitro and receptors on the surface of mammalian cells.

Mingjie Zhang - One of the best experts on this subject based on the ideXlab platform.

  • Versatile Site-Selective Protein Reaction Guided by WW Domain-Peptide Motif Interaction.
    Bioconjugate chemistry, 2017
    Co-Authors: Miao Liu, Mingjie Zhang, Jiang Xia
    Abstract:

    A short, flexible, and unstructured peptide tag that has versatile and facile use in protein labeling applications is highly desirable. Here, we report an 11-residue peptide tag with an internal cysteine (a W-tag, derived from a Comm PY peptide motif that is known to bind with Nedd4 WW3* domain) that can be installed at different regions of the target protein without compromising its covalent reactivity with the reactive label (a 35-residue synthetic Nedd4 WW3* domain derivative). This versatility is explained by the unique structural features of the reaction. NMR analysis reveals that both the W-tag peptide and reactive Nedd4 WW3* protein are unstructured before they encounter each other. The binding interaction of the two induces noticeable structural changes and promotes global folding. Consequently, the reactive cysteine residue at W-tag and the electrophilic Chloroacetyl Group at Nedd4 WW3* domain are positioned to be in close proximity, inducing an intermolecular covalent cross-linking. The covalent...

  • Versatile Site-Selective Protein Reaction Guided by WW Domain–Peptide Motif Interaction
    2017
    Co-Authors: Miao Liu, Mingjie Zhang, Jiang Xia
    Abstract:

    A short, flexible, and unstructured peptide tag that has versatile and facile use in protein labeling applications is highly desirable. Here, we report an 11-residue peptide tag with an internal cysteine (a W-tag, derived from a Comm PY peptide motif that is known to bind with Nedd4 WW3* domain) that can be installed at different regions of the target protein without compromising its covalent reactivity with the reactive label (a 35-residue synthetic Nedd4 WW3* domain derivative). This versatility is explained by the unique structural features of the reaction. NMR analysis reveals that both the W-tag peptide and reactive Nedd4 WW3* protein are unstructured before they encounter each other. The binding interaction of the two induces noticeable structural changes and promotes global folding. Consequently, the reactive cysteine residue at W-tag and the electrophilic Chloroacetyl Group at Nedd4 WW3* domain are positioned to be in close proximity, inducing an intermolecular covalent cross-linking. The covalent linkage in turn stabilizes the folding of the protein complex. This unique multistep mechanism renders this labeling reaction amenable to different sites of the proteins of interest: installation of the tag at N- and C-termini, in the flexible linker region, in the loop region, and the extracellular terminus of target proteins exhibited comparable reactivity. This work therefore represents the first proximity-induced cysteine reaction based on the unique binding features of WW domains that demonstrates unprecedented versatility