1XH0

Structure of the N298S variant of human pancreatic alpha-amylase complexed with acarbose

1XH0 の概要

• エントリーDOI: 10.2210/pdb1xh0/pdb
• 関連するPDBエントリー: 1XGZ 1XH1 1XH2
• 分子名称: Alpha-amylase, pancreatic, 2-acetamido-2-deoxy-beta-D-glucopyranose, ACARBOSE DERIVED HEXASACCHARIDE, ... (4 entities in total)
• 機能のキーワード: chloride amylase enzyme acarbose inhibitor, hydrolase
• 由来する生物種: Homo sapiens (human)
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 57095.38

構造登録者

Maurus, R.,Begum, A.,Kuo, H.H.,Racaza, A.,Numao, S.,Overall, C.M.,Withers, S.G.,Brayer, G.D. (登録日: 2004-09-17, 公開日: 2005-05-24, 最終更新日: 2024-10-30)

主引用文献

Maurus, R.,Begum, A.,Kuo, H.H.,Racaza, A.,Numao, S.,Andersen, C.,Tams, J.W.,Vind, J.,Overall, C.M.,Withers, S.G.,Brayer, G.D.
Structural and mechanistic studies of chloride induced activation of human pancreatic alpha-amylase
PROTEIN SCI., 14:743-755, 2005

PubMed Abstract: The mechanism of allosteric activation of alpha-amylase by chloride has been studied through structural and kinetic experiments focusing on the chloride-dependent N298S variant of human pancreatic alpha-amylase (HPA) and a chloride-independent TAKA-amylase. Kinetic analysis of the HPA variant clearly demonstrates the pronounced activating effect of chloride ion binding on reaction rates and its effect on the pH-dependence of catalysis. Structural alterations observed in the N298S variant upon chloride ion binding suggest that the chloride ion plays a variety of roles that serve to promote catalysis. One of these is having a strong influence on the positioning of the acid/base catalyst residue E233. Absence of chloride ion results in multiple conformations for this residue and unexpected enzymatic products. Chloride ion and N298 also appear to stabilize a helical region of polypeptide chain from which projects the flexible substrate binding loop unique to chloride-dependent alpha-amylases. This structural feature also serves to properly orient the catalytically essential residue D300. Comparative analyses show that the chloride-independent alpha-amylases compensate for the absence of bound chloride by substituting a hydrophobic core, altering the manner in which substrate interactions are made and shifting the placement of N298. These evolutionary differences presumably arise in response to alternative operating environments or the advantage gained in a particular product profile. Attempts to engineer chloride-dependence into the chloride-independent TAKA-amylase point out the complexity of this system, and the fact that a multitude of factors play a role in binding chloride ion in the chloride-dependent alpha-amylases.

PubMed: 15722449
DOI: 10.1110/ps.041079305

実験手法

X-RAY DIFFRACTION (2 Å)