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.
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 alpha-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.
Chong
B E
BE
Department of Chemistry, University of Michigan, Ann Arbor 48109-1055, USA.
Wall
D B
DB
Lubman
D M
DM
Flynn
S J
SJ
D001426:Bacterial Proteins / Q000737:chemistry*; D002109:Caffeic Acids / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid; D003101:Collodion; D003373:Coumaric Acids / Q000737:chemistry; D003470:Culture Media; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000737:chemistry*; D008970:Molecular Weight; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D001426:Bacterial Proteins; D002109:Caffeic Acids; D003373:Coumaric Acids; D003470:Culture Media; C007175:alpha-cyano-4-hydroxycinnamate; C073734:sinapinic acid; D003101:Collodion; C040048:caffeic acid