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Rapid profiling of E. coli proteins up to 500 kDa from whole cell lysates using matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry
Author(s) -
Chong Bathsheba E.,
Wall Daniel B.,
Lubman David M.,
Flynn Shan J.
Publication year - 1997
Publication title -
rapid communications in mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.528
H-Index - 136
eISSN - 1097-0231
pISSN - 0951-4198
DOI - 10.1002/(sici)1097-0231(199711)11:17<1900::aid-rcm95>3.0.co;2-k
Subject(s) - chemistry , chromatography , mass spectrometry , matrix assisted laser desorption/ionization , desorption , surface enhanced laser desorption/ionization , guanidine , protein mass spectrometry , time of flight mass spectrometry , escherichia coli , biochemistry , ionization , tandem mass spectrometry , organic chemistry , ion , adsorption , gene
Matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry was used to rapidly detect and profile large proteins from Escherichia coli whole cell lysates in the mass range 25–500 kDa. The bacterial samples were treated with guanidine hydrochloride and Triton X‐100 to disrupt and solubilize the large inner membrane proteins. A sample preparation involving a nitrocellulose polymer film, and α‐cyano‐4‐hydroxycinnamic acid, sinapinic acid or caffeic acid as matrix was utilized to rapidly monitor the presence of induced and repressed protein synthesis in response to l ‐arabinose catabolism in E. coli cells. The results were compared to those of 1‐D or 2‐D gel electrophoresis. © 1997 John Wiley & Sons, Ltd.