1LCP

BOVINE LENS LEUCINE AMINOPEPTIDASE COMPLEXED WITH L-LEUCINE PHOSPHONIC ACID

1LCP の概要

• エントリーDOI: 10.2210/pdb1lcp/pdb
• 分子名称: LEUCINE AMINOPEPTIDASE, ZINC ION, LEUCINE PHOSPHONIC ACID, ... (5 entities in total)
• 機能のキーワード: hydrolase (alpha-aminoacylpeptide)
• 由来する生物種: Bos taurus (cattle)
• 細胞内の位置: Cytoplasm: P00727
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 106773.49

構造登録者

Straeter, N.,Lipscomb, W.N. (登録日: 1995-05-12, 公開日: 1995-07-31, 最終更新日: 2024-02-14)

主引用文献

Strater, N.,Lipscomb, W.N.
Transition state analogue L-leucinephosphonic acid bound to bovine lens leucine aminopeptidase: X-ray structure at 1.65 A resolution in a new crystal form.
Biochemistry, 34:9200-9210, 1995

PubMed Abstract: The three-dimensional structure of bovine lens leucine aminopeptidase (blLAP) complexed with L-Leucinephosphonic acid (LeuP) has been determined by molecular replacement using the structure of native blLAP as a starting model. Cocrystallization of the enzyme with the inhibitor yielded a new crystal form of space group P321 which has cell dimensions a = 130.4 A and c = 125.4 A. Refinement of the model against data from 7.0 to 1.65 A resolution resulted in a final structure with a crystallographic residual of 0.160 (R(free) = 0.191). The N-terminal amino group of LeuP is coordinated to Zn-489, one phosphoryl oxygen atom bridges both metal ions, and another phosphoryl oxygen atom is coordinated to Zn-488. The side chain of Arg-336 interacts with the inhibitor via three water molecules. LeuP resembles the presumed tetrahedral gem-diolate transition state after direct attack of a water or hydroxide ion nucleophile on the scissile peptide bond. On the basis of the LeuP binding mode and the previous structural and biochemical data, three plausible reaction pathways are evaluated. The two-metal ion mechanisms discussed herein share as common features a metal-bound hydroxide ion nucleophile and polarization of the carbonyl group by the zinc ions. Possible catalytic roles of Arg-336 and Lys-262 in the direct or indirect (through H2O) protonation of the leaving group, in the stabilization of a zinc-bound OH- nucleophile and in the stabilization of the negatively charged intermediate, are discussed. A site 3 metal ion approximately 12 A away from the active site 2 zinc ion probably serves a structural role.

PubMed: 7619821
DOI: 10.1021/bi00028a033

実験手法

X-RAY DIFFRACTION (1.65 Å)