DOI: 10.1073/pnas.1715127114
论文题名: Cholesterol-binding site of the influenza M2 protein in lipid bilayers from solid-state NMR
作者: Elkins M.R. ; Williams J.K. ; Gelenter M.D. ; Dai P. ; Kwon B. ; Sergeyev I.V. ; Pentelute B.L. ; Hong M.
刊名: Proceedings of the National Academy of Sciences of the United States of America
ISSN: 0027-8424
出版年: 2017
卷: 114, 期: 49 起始页码: 12946
结束页码: 12951
语种: 英语
英文关键词: 19F-NMR
; Deuterium NMR
; Docking
; Membrane proteins
; Membrane scission
Scopus关键词: cholesterol
; membrane protein
; methyl group
; protein M2
; tetramer
; cholesterol
; M2 protein, Influenza A virus
; matrix protein
; Article
; binding site
; carboxy terminal sequence
; deuteron nuclear magnetic resonance
; hydrophobicity
; Influenza virus
; molecular interaction
; molecular recognition
; molecular stability
; nonhuman
; nuclear magnetic resonance spectroscopy
; phospholipid bilayer
; phosphorus nuclear magnetic resonance
; priority journal
; protein motif
; virus release
; alpha helix
; chemistry
; Influenza A virus
; lipid bilayer
; molecular docking
; nuclear magnetic resonance
; protein domain
; ultrastructure
; Binding Sites
; Cholesterol
; Influenza A virus
; Lipid Bilayers
; Molecular Docking Simulation
; Nuclear Magnetic Resonance, Biomolecular
; Protein Conformation, alpha-Helical
; Protein Domains
; Viral Matrix Proteins
英文摘要: The influenza M2 protein not only forms a proton channel but also mediates membrane scission in a cholesterol-dependent manner to cause virus budding and release. The atomic interaction of cholesterol with M2, as with most eukaryotic membrane proteins, has long been elusive. We have now determined the cholesterol-binding site of the M2 protein in phospholipid bilayers using solid-state NMR spectroscopy. Chain-fluorinated cholesterol was used to measure cholesterol proximity to M2 while sterol-deuterated cholesterol was used to measure bound-cholesterol orientation in lipid bilayers. Carbon–fluorine distance measurements show that at a cholesterol concentration of 17 mol%, two cholesterol molecules bind each M2 tetramer. Cholesterol binds the C-terminal transmembrane (TM) residues, near an amphipathic helix, without requiring a cholesterol recognition sequence motif. Deuterium NMR spectra indicate that bound cholesterol is approximately parallel to the bilayer normal, with the rough face of the sterol rings apposed to methyl-rich TM residues. The distance- and orientation-restrained cholesterol-binding site structure shows that cholesterol is stabilized by hydrophobic interactions with the TM helix and polar and aromatic interactions with neighboring amphipathic helices. At the 1:2 binding stoichiometry, lipid 31P spectra show an isotropic peak indicative of high membrane curvature. This M2–cholesterol complex structure, together with previously observed M2 localization at phase boundaries, suggests that cholesterol mediates M2 clustering to the neck of the budding virus to cause the necessary curvature for membrane scission. The solid-state NMR approach developed here is generally applicable for elucidating the structural basis of cholesterol’s effects on membrane protein function. © 2017, National Academy of Sciences. All rights reserved. Citation statistics:
资源类型: 期刊论文
标识符: http://119.78.100.158/handle/2HF3EXSE/163768
Appears in Collections: 气候变化与战略
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作者单位: Elkins, M.R., Department of Chemistry, Massachusetts Institute of Technology, Cambridge, MA 02139, United States; Williams, J.K., Department of Chemistry, Massachusetts Institute of Technology, Cambridge, MA 02139, United States; Gelenter, M.D., Department of Chemistry, Massachusetts Institute of Technology, Cambridge, MA 02139, United States; Dai, P., Department of Chemistry, Massachusetts Institute of Technology, Cambridge, MA 02139, United States; Kwon, B., Department of Chemistry, Massachusetts Institute of Technology, Cambridge, MA 02139, United States; Sergeyev, I.V., Bruker Biospin, Billerica, MA 01821, United States; Pentelute, B.L., Department of Chemistry, Massachusetts Institute of Technology, Cambridge, MA 02139, United States; Hong, M., Department of Chemistry, Massachusetts Institute of Technology, Cambridge, MA 02139, United States
Recommended Citation:
Elkins M.R.,Williams J.K.,Gelenter M.D.,et al. Cholesterol-binding site of the influenza M2 protein in lipid bilayers from solid-state NMR[J]. Proceedings of the National Academy of Sciences of the United States of America,2017-01-01,114(49)