Research article summary (published 31 Aug 2009):

The role of Tyr71 in Streptomyces trypsin on the recognition mechanism of structural protein substrates.

Full Abstract

Studies of substrate recognition by serine proteases have focused on specificities at the primary S1-Sn sites, but topological specificities (i.e. recognition at distinct three-dimensional structural motifs) have not been established. This is the first report to identify the key amino acid residue conferring topological specificity. A serine protease from Streptomyces omiyaensis (SOT), which is a trypsin-like enzyme, was chosen as a model enzyme to clarify the recognition mechanism of structural protein substrates in serine proteases. We have found previously that the topological specificities of SOT and S. griseus trypsin (SGT) for high molecular mass substrates differ greatly, even though the enzymes have similar primary structures. In this study, we constructed chimeras between SOT and SGT using an in vivo DNA shuffling system and several mutants to identify the key residues involved in topological specificities. By comparing the substrate specificities of chimeras and mutants, we found that residue 71 of SOT, which is separate from the catalytic triad, contributes to the topological specificity. Using site-directed mutagenesis, residue 71 of SOT was also found to be crucial for catalytic efficiency and enzyme conformation.

Author information

Author/s: Uesugi, Yoshiko (Y); Usuki, Hirokazu (H); Iwabuchi, Masaki (M); Hatanaka, Tadashi (T);

Affiliation: Research Institute for Biological Sciences, Okayama, Japan.

Journal and publication information

Publication Type: In Vitro; Journal Article; Research Support, Non-U.S. Gov't

Journal: The FEBS journal (FEBS J), published in England. (Language: eng)

Reference: 2009-Oct; vol 276 (issue 19) : pp 5634-46

Dates: Created 2009/09/22; Completed 2009/10/22;

PMID: 19725878, status: MEDLINE (last retrieval date: 10/22/2009, IMS Date: )

Sourced from the National Library of Medicine. Abstract text and other information may be subject to copyright.

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MeSH headings (categories)

This article was linked to the MESH Headings shown below.

Amino Acid Sequence Amino Acid Substitution Animals Base Sequence Catalytic Domain - genetics Cattle Collagen Type I - metabolism Collagen Type IV - metabolism DNA Primers - genetics Genes, Bacterial Humans Models, Molecular

Models, Molecular Molecular Sequence Data Mutagenesis, Site-Directed Protein Conformation Recombinant Fusion Proteins - chemistry; genetics; metabolism Sequence Homology, Amino Acid Species Specificity Streptomyces - enzymology; genetics Streptomyces griseus - enzymology; genetics Substrate Specificity Trypsin - chemistry; genetics; metabolism Tyrosine - chemistry

Associated Chemicals: Collagen Type I (0) ; Collagen Type IV (0) ; DNA Primers (0) ; Recombinant Fusion Proteins (0) ; Tyrosine (55520-40-6) ; Trypsin (EC 3.4.21.4)

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