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PMID: 9238014 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Complete resolution of the solid-state NMR spectrum of a uniformly 15N-labeled membrane protein in phospholipid bilayers.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 94 ·No. 16 ·1997-08-05 ·Pages 8551-6

Marassi FM, Ramamoorthy A, Opella SJ

Abstract

Complete resolution of the amide resonances in a three-dimensional solid-state NMR correlation spectrum of a uniformly 15N-labeled membrane protein in oriented phospholipid bilayers is demonstrated. The three orientationally dependent frequencies, 1H chemical shift, 1H-15N dipolar coupling, and 15N chemical shift, associated with each amide resonance are responsible for resolution among resonances and provide sufficient angular restrictions for protein structure determination. Because the protein is completely immobilized by the phospholipids on the relevant NMR time scales (10 kHz), the linewidths will not degrade in the spectra of larger proteins. Therefore, these results demonstrate that solid-state NMR experiments can overcome the correlation time problem and extend the range of proteins that can have their structures determined by NMR spectroscopy to include uniformly 15N-labeled membrane proteins in phospholipid bilayers.

MeSH Terms
Amino Acid Sequence Escherichia coli Lipid Bilayers/chemistry Magnetic Resonance Spectroscopy Membrane Proteins/chemistry Molecular Sequence Data Phospholipids/chemistry Protein Binding
Chemicals
Lipid Bilayers Membrane Proteins Phospholipids
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Marassi F M
Department of Chemistry, University of Pennsylvania, Philadelphia, PA 19104, USA.
Ramamoorthy A
Opella S J
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.94.16.8551
Published
1997-08-05
Pages
8551-6
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC23006
Subset
IM
Grants
NIGMS NIH HHS · R01 GM029754 · United States
NIAID NIH HHS · R01 AI20770 · United States
NIGMS NIH HHS · R01 GM29754 · United States
NIGMS NIH HHS · R37 GM24266 · United States
External Links
PubMed source DOI: 10.1073/pnas.94.16.8551 PMC Full Text View journal details
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