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
Rok vydání: 1999
Předmět:
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