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Substrate specificity of microbial transglutaminase as revealed by three-dimensional docking simulation and mutagenesis.

ABSTRACT: Transglutaminases (TGases) are used in fields such as food and pharmaceuticals. Unlike other TGases, microbial transglutaminase (MTG) activity is Ca(2+)-independent, broadening its application. Here, a three-dimensional docking model of MTG binding to a peptide substrate, CBZ-Gln-Gly, was simulated. The data reveal CBZ-Gln-Gly to be stretched along the MTG active site cleft with hydrophobic and/or aromatic residues interacting directly with the substrate. Moreover, an oxyanion binding site for TGase activity may be constructed from the amide groups of Cys64 and/or Val65. Alanine mutagenesis verified the simulated binding region and indicated that large molecules can be widely recognized on the MTG cleft.

SUBMITTER: Tagami U

PROVIDER: S-EPMC2777024 | biostudies-literature | 2009 Dec

REPOSITORIES: biostudies-literature

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Substrate specificity of microbial transglutaminase as revealed by three-dimensional docking simulation and mutagenesis.

Tagami Uno U Shimba Nobuhisa N Nakamura Mina M Yokoyama Kei-Ichi K Suzuki Ei-Ichiro E Hirokawa Takatsugu T

Protein engineering, design & selection : PEDS 20091022 12

Transglutaminases (TGases) are used in fields such as food and pharmaceuticals. Unlike other TGases, microbial transglutaminase (MTG) activity is Ca(2+)-independent, broadening its application. Here, a three-dimensional docking model of MTG binding to a peptide substrate, CBZ-Gln-Gly, was simulated. The data reveal CBZ-Gln-Gly to be stretched along the MTG active site cleft with hydrophobic and/or aromatic residues interacting directly with the substrate. Moreover, an oxyanion binding site for T ...[more]

PMID: 19850674

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