Toward the bilayer proteome, electrospray ionization-mass spectrometry of large, intact transmembrane proteins
Autor: | Jenny C. Lee, H R Kaback, Gilbert G. Privé, J. le Coutre, Christian K Engel, Kym F. Faull, Julian P. Whitelegge |
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Rok vydání: | 1999 |
Předmět: |
Lactose permease
Time Factors Multidisciplinary Monosaccharide Transport Proteins Symporters biology Chemistry Membrane transport protein Escherichia coli Proteins Electrospray ionization Membrane Proteins Membrane Transport Proteins Biological Sciences Mass spectrometry Mass Spectrometry Transmembrane protein Biochemistry Membrane protein Proteome Chromatography Gel Escherichia coli biology.protein Chromatography High Pressure Liquid |
Zdroj: | Proceedings of the National Academy of Sciences. 96:10695-10698 |
ISSN: | 1091-6490 0027-8424 |
DOI: | 10.1073/pnas.96.19.10695 |
Popis: | Genes encoding membrane proteins comprise a substantial proportion of genomes sequenced to date, but ability to perform structural studies on this portion of the proteome is limited. Electrospray ionization-MS (ESI-MS) of an intact protein generates a profile defining the native covalent state of the gene product and its heterogeneity. Here we apply ESI-MS technology with accuracy exceeding 0.01% to a hydrophobic membrane protein with 12-transmembrane α-helices, the full-length lactose permease from Escherichia coli . Furthermore, ESI-MS is used to titrate reactive thiols with N -ethylmaleimide. Treatment of the native protein solubilized in detergent micelles reveals only two reactive thiols, and both are protected by a substrate analog. |
Databáze: | OpenAIRE |
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