In situ structural characterization of a recombinant protein in native Escherichia coli membranes with solid-state magic-angle-spinning NMR

Riqiang Fu, Xingsheng Wang, Conggang Li, Adriana N. Santiago-Miranda, Gary J. Pielak, Fang Tian

Research output: Contribution to journalArticle

53 Citations (Scopus)

Abstract

The feasibility of using solid-state magic-angle-spinning NMR spectroscopy for in situ structural characterization of the LR11 (sorLA) transmembrane domain (TM) in native Escherichia coli membranes is presented. LR11 interacts with the human amyloid precursor protein (APP), a central player in the pathology of Alzheimer's disease. The background signals from E. coli lipids and membrane proteins had only minor effects on the LR11 TM resonances. Approximately 50% of the LR11 TM residues were assigned by using 13C PARIS data. These assignments allowed comparisons of the secondary structure of the LR11 TM in native membrane environments and commonly used membrane mimics (e.g., micelles). In situ spectroscopy bypasses several obstacles in the preparation of membrane proteins for structural analysis and offers the opportunity to investigate how membrane heterogeneity, bilayer asymmetry, chemical gradients, and macromolecular crowding affect the protein structure.

Original languageEnglish (US)
Pages (from-to)12370-12373
Number of pages4
JournalJournal of the American Chemical Society
Volume133
Issue number32
DOIs
StatePublished - Aug 17 2011

Fingerprint

Recombinant proteins
Magic angle spinning
Recombinant Proteins
Escherichia coli
Nuclear magnetic resonance
Membranes
Proteins
Membrane Proteins
Crowding
Amyloid beta-Protein Precursor
Escherichia coli Proteins
Micelles
Pathology
Structural analysis
Lipids
Nuclear magnetic resonance spectroscopy
Spectrum Analysis
Alzheimer Disease
Magnetic Resonance Spectroscopy
Spectroscopy

All Science Journal Classification (ASJC) codes

  • Catalysis
  • Chemistry(all)
  • Biochemistry
  • Colloid and Surface Chemistry

Cite this

Fu, Riqiang ; Wang, Xingsheng ; Li, Conggang ; Santiago-Miranda, Adriana N. ; Pielak, Gary J. ; Tian, Fang. / In situ structural characterization of a recombinant protein in native Escherichia coli membranes with solid-state magic-angle-spinning NMR. In: Journal of the American Chemical Society. 2011 ; Vol. 133, No. 32. pp. 12370-12373.
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In situ structural characterization of a recombinant protein in native Escherichia coli membranes with solid-state magic-angle-spinning NMR. / Fu, Riqiang; Wang, Xingsheng; Li, Conggang; Santiago-Miranda, Adriana N.; Pielak, Gary J.; Tian, Fang.

In: Journal of the American Chemical Society, Vol. 133, No. 32, 17.08.2011, p. 12370-12373.

Research output: Contribution to journalArticle

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