1AND

ANIONIC TRYPSIN MUTANT WITH ARG 96 REPLACED BY HIS

1AND の概要

• エントリーDOI: 10.2210/pdb1and/pdb
• 分子名称: ANIONIC TRYPSIN, COPPER (II) ION, BENZAMIDINE, ... (4 entities in total)
• 機能のキーワード: trypsin, anionic, serine protease, hydrolase
• 由来する生物種: Rattus rattus (black rat)
• 細胞内の位置: Secreted, extracellular space: P00763
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 23979.49

構造登録者

Fletterick, R.J.,Mcgrath, M.E. (登録日: 1994-12-21, 公開日: 1997-04-01, 最終更新日: 2024-11-20)

主引用文献

McGrath, M.E.,Haymore, B.L.,Summers, N.L.,Craik, C.S.,Fletterick, R.J.
Structure of an engineered, metal-actuated switch in trypsin.
Biochemistry, 32:1914-1919, 1993

PubMed Abstract: The X-ray crystal structure of the copper complex of the rat trypsin mutant Arg96 to His96 (trypsin R96H) has been determined in order to ascertain the nature of the engineered metal-binding site and to understand the structural basis for the metal-induced enzymatic inhibition. In the structure, the catalytically essential His57 residue is reoriented out of the active-site pocket and forms a chelating, metal-binding site with residue His96. The copper is bound to the N epsilon 2 atoms of both histidine residues with Cu-N epsilon 2 = 2.2 A and N epsilon 2-Cu-N epsilon 2 = 89 degrees. The metal is clearly bound to a third ligand leading to a distorted square planar geometry at Cu. The X-ray results do not unambiguously yield the identity of this third ligand, but chemical data suggest that it is a deprotonated, chelating Tris molecule which was used as a carrier to solubilize the copper in alkaline solution (pH 8.0). Upon reorientation of His57, a unique water molecule moves into the active site and engages in hydrogen-bonding with Asp102-O delta 2 and His57-N delta 1. Except for small movements of the peptide backbone near His96, the remainder of the trypsin molecule is isostructural with the native enzyme. These data support the notion that the effective inhibition of catalytic activity by metal ions observed in trypsin R96H is indeed caused by a specific and reversible reorganization of the active site in the enzyme.

PubMed: 8448149
DOI: 10.1021/bi00059a005

実験手法

X-RAY DIFFRACTION (2.3 Å)