Identification of proteins from Escherichia coli using two-dimensional semi-preparative electrophoresis and mass spectrometry.
Escherichia coli is a gram-negative bacterium that causes sepsis and infections of the nervous system, and the digestive and urinary tracts. The availability of the complete nucleotide sequence encoding the E. coli K-12 genome has made this organism an excellent model for proteomic studies. Semi-preparative two-dimensional electrophoresis, including liquid phase isoelectric focusing (IEF), one-dimensional sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis (PAGE) and gel elution, have for the first time been used in combination with matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOFMS), electrospray tandem mass spectrometry and database searching for rapid separation of proteins from a uropathogenic strain of E. coli. The identity of 30 proteins, including the membrane protein nmpC, was obtained using this approach.
Gustafsson
E
E
Institute of Medical Biochemistry, Göteborg University, Box 440, SE-405 30 Göteborg, Sweden.
Thorén
K
K
Larsson
T
T
Davidsson
P
P
Karlsson
K A
KA
Nilsson
C L
CL
D001426:Bacterial Proteins / Q000032:analysis* / Q000302:isolation & purification; D002458:Cell Fractionation; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D004926:Escherichia coli; D007525:Isoelectric Focusing; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
D001426:Bacterial Proteins