1OSS

T190P STREPTOMYCES GRISEUS TRYPSIN IN COMPLEX WITH BENZAMIDINE

1OSS の概要

• エントリーDOI: 10.2210/pdb1oss/pdb
• 関連するPDBエントリー: 1OS8 1SGT
• 分子名称: trypsin, SULFATE ION, CALCIUM ION, ... (5 entities in total)
• 機能のキーワード: trypsin, serine protease, mutant, hydrolase
• 由来する生物種: Streptomyces griseus
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 23471.15

構造登録者

Page, M.J.,Wong, S.L.,Hewitt, J.,Strynadka, N.C.,MacGillivray, R.T. (登録日: 2003-03-20, 公開日: 2003-08-19, 最終更新日: 2024-11-06)

主引用文献

Page, M.J.,Wong, S.L.,Hewitt, J.,Strynadka, N.C.,MacGillivray, R.T.
Engineering the Primary Substrate Specificity of Streptomyces griseus Trypsin.
Biochemistry, 42:9060-9066, 2003

PubMed Abstract: 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.

PubMed: 12885239
DOI: 10.1021/bi0344230

実験手法

X-RAY DIFFRACTION (1.93 Å)