1IY5

Solution structure of wild type OMSVP3

1IY5 の概要

• エントリーDOI: 10.2210/pdb1iy5/pdb
• 関連するPDBエントリー: 1IY6
• NMR情報: BMRB: 5469
• 分子名称: OMSVP3 (1 entity in total)
• 機能のキーワード: solution structure, csh motif, omsvp3, ovomucoid third domain, protease inhibitor, hydrolase
• 由来する生物種: Lophura nycthemera (silver pheasant)
• 細胞内の位置: Secreted: P67954
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 5860.61

構造登録者

Hemmi, H.,Kumazaki, T.,Yamazaki, T.,Kojima, S.,Yoshida, T.,Kyogoku, Y.,Katsu, M.,Yokosawa, H.,Miura, K.,Kobayashi, Y. (登録日: 2002-07-23, 公開日: 2003-03-11, 最終更新日: 2024-10-30)

主引用文献

Hemmi, H.,Kumazaki, T.,Yamazaki, T.,Kojima, S.,Yoshida, T.,Kyogoku, Y.,Katsu, M.,Shinohara, F.,Yokosawa, H.,Miura, K.,Kobayashi, Y.
Inhibitory Specificity Change of Ovomucoid Third Domain of the Silver Pheasant upon Introduction of an Engineered Cys14-Cys39 Bond
BIOCHEMISTRY, 42:2524-2534, 2003

PubMed Abstract: The ovomucoid third domain from silver pheasant (OMSVP3), a typical Kazal-type inhibitor, strongly inhibits different serine proteases of various specificities, i.e., chymotrypsin, Streptomyces griseus protease, subtilisin, and elastase. Structural studies have suggested that conformational flexibility in the reactive site loop of the free inhibitor may be related to broad specificity of the ovomucoid. On the basis of the structural homology between OMSVP3 and ascidian trypsin inhibitor (ATI), which has a cystine-stabilized alpha-helical (CSH) motif in the sequence, we prepared the disulfide variant of OMSVP3, introducing an engineered disulfide bond between positions 14 and 39 near the reactive site (Met18-Glu19) by site-directed mutagenesis. The disulfide variant P14C/N39C retained potent inhibitory activities toward alpha-chymotrypsin (CHT) and S. griseus proteases A and B (SGPA and SGPB), while this variant lost most of its inhibitory activity toward porcine pancreatic elastase (PPE). We determined the solution structure of P14C/N39C, as well as that of wild-type OMSVP3, by two-dimensional nuclear magnetic resonance (2D NMR) methods and compared their structures to elucidate the structural basis of the inhibitory specificity change. For the molecular core consisting of a central alpha-helix and a three-stranded antiparallel beta-sheet, essentially no structural difference was detected between the two (pairwise rmsd value = 0.41 A). In contrast to this, a significant difference was detected in the loop from Cys8 to Thr17, where in P14C/N39C it has drawn approximately 4 A nearer the central helix to form the engineered Cys14-Cys39 bond. Concomitantly, the Tyr11-Pro12 cis-peptide linkage, which is highly conserved in ovomucoid third domains, was isomerized to the trans configuration. Such structural change in the loop near the reactive site may possibly affect the inhibitory specificity of P14C/N39C for the corresponding proteases.

PubMed: 12614146
DOI: 10.1021/bi026727c

実験手法

SOLUTION NMR