3H8F

High pH native structure of leucine aminopeptidase from Pseudomonas putida

3H8F の概要

• エントリーDOI: 10.2210/pdb3h8f/pdb
• 関連するPDBエントリー: 1BLL 1GYT 3H8E 3H8G
• 分子名称: Cytosol aminopeptidase, ZINC ION, MANGANESE (II) ION, ... (6 entities in total)
• 機能のキーワード: aminopeptidase, hydrolase, manganese, metal-binding, protease
• 由来する生物種: Pseudomonas putida
• 細胞内の位置: Cytoplasm : O86436
• タンパク質・核酸の鎖数: 6
• 化学式量合計: 316510.84

構造登録者

Kale, A.,Dijkstra, B.W.,Sonke, T.,Thunnissen, A.M.W.H. (登録日: 2009-04-29, 公開日: 2010-04-14, 最終更新日: 2024-02-21)

主引用文献

Kale, A.,Pijning, T.,Sonke, T.,Dijkstra, B.W.,Thunnissen, A.M.
Crystal structure of the leucine aminopeptidase from Pseudomonas putida reveals the molecular basis for its enantioselectivity and broad substrate specificity.
J.Mol.Biol., 398:703-714, 2010

PubMed Abstract: The zinc-dependent leucine aminopeptidase from Pseudomonas putida (ppLAP) is an important enzyme for the industrial production of enantiomerically pure amino acids. To provide a better understanding of its structure-function relationships, the enzyme was studied by X-ray crystallography. Crystal structures of native ppLAP at pH 9.5 and pH 5.2, and in complex with the inhibitor bestatin, show that the overall folding and hexameric organization of ppLAP are very similar to those of the closely related di-zinc leucine aminopeptidases (LAPs) from bovine lens and Escherichia coli. At pH 9.5, the active site contains two metal ions, one identified as Mn(2+) or Zn(2+) (site 1), and the other as Zn(2+) (site 2). By using a metal-dependent activity assay it was shown that site 1 in heterologously expressed ppLAP is occupied mainly by Mn(2+). Moreover, it was shown that Mn(2+) has a significant activation effect when bound to site 1 of ppLAP. At pH 5.2, the active site of ppLAP is highly disordered and the two metal ions are absent, most probably due to full protonation of one of the metal-interacting residues, Lys267, explaining why ppLAP is inactive at low pH. A structural comparison of the ppLAP-bestatin complex with inhibitor-bound complexes of bovine lens LAP, along with substrate modelling, gave clear and new insights into its substrate specificity and high level of enantioselectivity.

PubMed: 20359484
DOI: 10.1016/j.jmb.2010.03.042

実験手法

X-RAY DIFFRACTION (2.2 Å)