Crystal structure of peptide-bound neprilysin reveals key binding interactions

Research output: Contribution to journalArticle

Abstract

Neprilysin (NEP) is a promiscuous zinc metalloprotease with broad substrate specificity and cleaves a remarkable diversity of substrates through endopeptidase action. Two of these – amyloid-β and natriuretic peptides – implicate the enzyme in both Alzheimer’s disease and cardiovascular disease, respectively. Here, we report the creation of a catalytically inactive NEP (E584D) to determine the first peptide-bound crystal structure at 2.6 Å resolution. The structure reveals key interactions involved in substrate binding which we have identified to be conserved in other known zinc metalloproteases. In addition, the structure provides evidence for a potential exosite within the central cavity that may play a critical role in substrate positioning. Together, these results contribute to our understanding of the molecular function of NEP.

Original languageEnglish
Number of pages10
JournalFEBS Letters
Early online date12 Sep 2019
DOIs
Publication statusE-pub ahead of print - 12 Sep 2019

Keywords

  • crystallography
  • NEP
  • neprilysin
  • neutral endopeptidase
  • peptide-bound
  • protein structure
  • zinc metalloprotease

ASJC Scopus subject areas

  • Biophysics
  • Structural Biology
  • Biochemistry
  • Molecular Biology
  • Genetics
  • Cell Biology

Cite this

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title = "Crystal structure of peptide-bound neprilysin reveals key binding interactions",
abstract = "Neprilysin (NEP) is a promiscuous zinc metalloprotease with broad substrate specificity and cleaves a remarkable diversity of substrates through endopeptidase action. Two of these – amyloid-β and natriuretic peptides – implicate the enzyme in both Alzheimer’s disease and cardiovascular disease, respectively. Here, we report the creation of a catalytically inactive NEP (E584D) to determine the first peptide-bound crystal structure at 2.6 {\AA} resolution. The structure reveals key interactions involved in substrate binding which we have identified to be conserved in other known zinc metalloproteases. In addition, the structure provides evidence for a potential exosite within the central cavity that may play a critical role in substrate positioning. Together, these results contribute to our understanding of the molecular function of NEP.",
keywords = "crystallography, NEP, neprilysin, neutral endopeptidase, peptide-bound, protein structure, zinc metalloprotease",
author = "Stephen Moss and Vasanta Subramanian and Acharya, {K. Ravi}",
note = "{\circledC} 2019 Federation of European Biochemical Societies.",
year = "2019",
month = "9",
day = "12",
doi = "10.1002/1873-3468.13602",
language = "English",
journal = "FEBS Letters",
issn = "0014-5793",
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T1 - Crystal structure of peptide-bound neprilysin reveals key binding interactions

AU - Moss, Stephen

AU - Subramanian, Vasanta

AU - Acharya, K. Ravi

N1 - © 2019 Federation of European Biochemical Societies.

PY - 2019/9/12

Y1 - 2019/9/12

N2 - Neprilysin (NEP) is a promiscuous zinc metalloprotease with broad substrate specificity and cleaves a remarkable diversity of substrates through endopeptidase action. Two of these – amyloid-β and natriuretic peptides – implicate the enzyme in both Alzheimer’s disease and cardiovascular disease, respectively. Here, we report the creation of a catalytically inactive NEP (E584D) to determine the first peptide-bound crystal structure at 2.6 Å resolution. The structure reveals key interactions involved in substrate binding which we have identified to be conserved in other known zinc metalloproteases. In addition, the structure provides evidence for a potential exosite within the central cavity that may play a critical role in substrate positioning. Together, these results contribute to our understanding of the molecular function of NEP.

AB - Neprilysin (NEP) is a promiscuous zinc metalloprotease with broad substrate specificity and cleaves a remarkable diversity of substrates through endopeptidase action. Two of these – amyloid-β and natriuretic peptides – implicate the enzyme in both Alzheimer’s disease and cardiovascular disease, respectively. Here, we report the creation of a catalytically inactive NEP (E584D) to determine the first peptide-bound crystal structure at 2.6 Å resolution. The structure reveals key interactions involved in substrate binding which we have identified to be conserved in other known zinc metalloproteases. In addition, the structure provides evidence for a potential exosite within the central cavity that may play a critical role in substrate positioning. Together, these results contribute to our understanding of the molecular function of NEP.

KW - crystallography

KW - NEP

KW - neprilysin

KW - neutral endopeptidase

KW - peptide-bound

KW - protein structure

KW - zinc metalloprotease

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DO - 10.1002/1873-3468.13602

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