Hydrophobic peptide mapping of clinically relevant heptathelical membrane proteins by capillary electrophoresis

Maoqing Dong, Robert P. Oda, Michael A. Strausbauch, Peter J. Wettstein, James P. Landers, Laurence J Miller

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

The structural investigation of G protein-coupled receptors has been hindered by the lack of techniques to effectively resolve the hydrophobic peptides obtained by chemical or proteolytic cleavage, as well as the minute amounts of protein typically isolated. We have developed a capillary electrophoresis method for efficient separation of hydrophobic peptides using a cyanogen bromide digest of bacteriorhodopsin as a model for these clinically important membrane proteins. This procedure includes (i) solubilization of the protein digest in acetic acid; and (ii) electrophoresis using an acetic acid-based buffer system augmented by acetonitrile and hexane sulfonic acid, in a Polybrene-coated fused silica capillary. The potential for detection sensitivity to be increased at least 100-fold by use of on-line solid-phase extraction on C18-silica is shown. This approach is potentially useful for peptide fingerprinting of sparse and extremely hydrophobic membrane receptors.

Original languageEnglish (US)
Pages (from-to)1767-1774
Number of pages8
JournalElectrophoresis
Volume18
Issue number10
DOIs
StatePublished - 1997

Fingerprint

Capillary electrophoresis
Peptide Mapping
Capillary Electrophoresis
Silicon Dioxide
Acetic Acid
Membrane Proteins
Hexadimethrine Bromide
Bacteriorhodopsins
Cyanogen Bromide
Peptides
Sulfonic Acids
Solid Phase Extraction
Hexanes
G-Protein-Coupled Receptors
Electrophoresis
Buffers
Proteins
Fused silica
Membranes
acetonitrile

Keywords

  • Bacteriorhodopsin
  • Capillary electrophoresis
  • Hydrophobic peptide mapping
  • Solid-phase extraction

ASJC Scopus subject areas

  • Clinical Biochemistry

Cite this

Hydrophobic peptide mapping of clinically relevant heptathelical membrane proteins by capillary electrophoresis. / Dong, Maoqing; Oda, Robert P.; Strausbauch, Michael A.; Wettstein, Peter J.; Landers, James P.; Miller, Laurence J.

In: Electrophoresis, Vol. 18, No. 10, 1997, p. 1767-1774.

Research output: Contribution to journalArticle

Dong, Maoqing ; Oda, Robert P. ; Strausbauch, Michael A. ; Wettstein, Peter J. ; Landers, James P. ; Miller, Laurence J. / Hydrophobic peptide mapping of clinically relevant heptathelical membrane proteins by capillary electrophoresis. In: Electrophoresis. 1997 ; Vol. 18, No. 10. pp. 1767-1774.
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