4CTH

Neprilysin variant G399V,G714K in complex with phosphoramidon

4CTH の概要

• エントリーDOI: 10.2210/pdb4cth/pdb
• 関連するBIRD辞書のPRD_ID: PRD_000638
• 分子名称: NEPRILYSIN, 2-acetamido-2-deoxy-beta-D-glucopyranose, ZINC ION, ... (6 entities in total)
• 機能のキーワード: transferase, amyloid beta, alzheimer's disease, enzyme engineering
• 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: 83423.82

構造登録者

Webster, C.I.,Burrell, M.,Olsson, L.,Fowler, S.B.,Digby, S.,Sandercock, A.,Snijder, A.,Tebbe, J.,Haupts, U.,Grudzinska, J.,Jermutus, L.,Andersson, C. (登録日: 2014-03-13, 公開日: 2014-08-13, 最終更新日: 2024-11-13)

主引用文献

Webster, C.I.,Burrell, M.,Olsson, L.,Fowler, S.B.,Digby, S.,Sandercock, A.,Snijder, A.,Tebbe, J.,Haupts, U.,Grudzinska, J.,Jermutus, L.,Andersson, C.
Engineering Neprilysin Activity and Specificity to Create a Novel Therapeutic for Alzheimer'S Disease.
Plos One, 9:04001-, 2014

PubMed Abstract: Neprilysin is a transmembrane zinc metallopeptidase that degrades a wide range of peptide substrates. It has received attention as a potential therapy for Alzheimer's disease due to its ability to degrade the peptide amyloid beta. However, its broad range of peptide substrates has the potential to limit its therapeutic use due to degradation of additional peptides substrates that tightly regulate many physiological processes. We sought to generate a soluble version of the ectodomain of neprilysin with improved activity and specificity towards amyloid beta as a potential therapeutic for Alzheimer's disease. Extensive amino acid substitutions were performed at positions surrounding the active site and inner surface of the enzyme and variants screened for activity on amyloid beta 1-40, 1-42 and a variety of other physiologically relevant peptides. We identified several mutations that modulated and improved both enzyme selectivity and intrinsic activity. Neprilysin variant G399V/G714K displayed an approximately 20-fold improved activity on amyloid beta 1-40 and up to a 3,200-fold reduction in activity on other peptides. Along with the altered peptide substrate specificity, the mutant enzyme produced a markedly altered series of amyloid beta cleavage products compared to the wild-type enzyme. Crystallisation of the mutant enzyme revealed that the amino acid substitutions result in alteration of the shape and size of the pocket containing the active site compared to the wild-type enzyme. The mutant enzyme offers the potential for the more efficient degradation of amyloid beta in vivo as a therapeutic for the treatment of Alzheimer's disease.

PubMed: 25089527
DOI: 10.1371/JOURNAL.PONE.0104001

実験手法

X-RAY DIFFRACTION (2.15 Å)