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'''T190P STREPTOMYCES GRISEUS TRYPSIN IN COMPLEX WITH BENZAMIDINE''' | '''T190P STREPTOMYCES GRISEUS TRYPSIN IN COMPLEX WITH BENZAMIDINE''' | ||
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[[Category: Strynadka, N C.]] | [[Category: Strynadka, N C.]] | ||
[[Category: Wong, S L.]] | [[Category: Wong, S L.]] | ||
[[Category: | [[Category: Mutant]] | ||
[[Category: | [[Category: Serine protease]] | ||
[[Category: | [[Category: Trypsin]] | ||
''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Sat May 3 04:14:25 2008'' | |||
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Revision as of 04:14, 3 May 2008
T190P STREPTOMYCES GRISEUS TRYPSIN IN COMPLEX WITH BENZAMIDINE
OverviewOverview
Streptomyces griseus trypsin (SGT) was chosen as a model scaffold for the development of serine proteases with enhanced substrate specificity. Recombinant SGT has been produced in a Bacillus subtilis expression system in a soluble active form and purified to homogeneity. The recombinant and native proteases have nearly identical enzymatic properties and structures. Four SGT mutants with alterations in the S1 substrate binding pocket (T190A, T190P, T190S, and T190V) were also expressed. The T190P mutant demonstrated the largest shift to a preference for Arg versus Lys in the P1 site. This was shown by a minor reduction in catalytic activity toward an Arg-containing substrate (k(cat) reduction of 25%). The crystal structures of the recombinant SGT and the T190P mutant in a complex with the inhibitor benzamidine were obtained at high resolution (approximately 1.9 A). The increase in P1 specificity, achieved with minimal effect on the catalytic efficiency, demonstrates that the T190P mutant is an ideal candidate for the design of additional substrate specificity engineered into the S2 to S4 binding pockets.
About this StructureAbout this Structure
1OSS is a Single protein structure of sequence from Streptomyces griseus. Full crystallographic information is available from OCA.
ReferenceReference
Engineering the primary substrate specificity of Streptomyces griseus trypsin., Page MJ, Wong SL, Hewitt J, Strynadka NC, MacGillivray RT, Biochemistry. 2003 Aug 5;42(30):9060-6. PMID:12885239 Page seeded by OCA on Sat May 3 04:14:25 2008