Evaluation of enzymatic digestion and liquid chromatography‐mass spectrometry peptide mapping of the integral membrane protein bacteriorhodopsin
- 19 September 2002
- journal article
- research article
- Published by Wiley in Electrophoresis
- Vol. 23 (18) , 3224-3232
- https://doi.org/10.1002/1522-2683(200209)23:18<3224::aid-elps3224>3.0.co;2-#
Abstract
A method for the complete peptide mapping of the model integral membrane protein bacteri-orhodopsin is demonstrated. Utilizing more effective enzymatic digestion, procedures with capillary liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS) and tandem mass spectrometry (MS/MS), all predicted tryptic digestion products were detected, as well as peptides from all previously reported post-translational modifications of bacteriorhodopsin. A significant contribution of chymotryptic-like digestion products was also observed. A characterization of the behavior of hydrophobic integral membrane peptides in a reversed-phase liquid chromatographic separation is also provided. The method reported here offers improved compatibility of the solubilizing reagents with both the chromatography and mass spectrometry, rendering it suitable for high-throughput proteomic applications.Keywords
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