Identification of bacterial proteins observed in MALDI TOF mass spectra from whole cells.
Characteristic ions in the MALDI TOF mass spectra from bacterial cells have been associated with four known proteins. The proteins, observed both from cells and in filtered cellular suspensions, were isolated by HPLC and identified on the basis of their mass spectra and their partial amino acid sequence, determined using the Edman method (10-15 residues). The acid resistance proteins HdeA and HdeB give rise to ions near m/z 9735 and 9060 in MALDI TOF mass spectra from cells and from extracts of both Escherichia coli 1090 and Shigella flexneri PHS-1059. However, the proteins associated with proteolytic cleavage by the peptidase Lep, rather than the precursor proteins, were observed, both using cells and from cellular extracts. A cold-shock protein, CspA, was associated with the ion near m/z 7643 from Pseudomonas aeruginosa. Similarly, a cold-acclimation protein, CapB, was identified as the source of the ion near m/z 7684 in P. putida. This last protein was homologous with a known CapB from P. fragi. While these experiments involved the detection of known or homologous proteins from typical bacteria, this same approach could also be applied to the detection of unique proteins or biomarker proteins associated with other bacteria of public health significance.
Holland
R D
RD
Division of Chemistry, Food and Drug Administration, Jefferson, Arkansas 72079, USA.
Duffy
C R
CR
Rafii
F
F
Sutherland
J B
JB
Heinze
T M
TM
Holder
C L
CL
Voorhees
K J
KJ
Lay
J O
JO
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000032:analysis* / Q000302:isolation & purification; D002352:Carrier Proteins / Q000032:analysis / Q000302:isolation & purification; D002851:Chromatography, High Pressure Liquid; D003080:Cold Temperature; D004926:Escherichia coli / Q000737:chemistry; D029968:Escherichia coli Proteins; D006360:Heat-Shock Proteins / Q000032:analysis / Q000302:isolation & purification; D008969:Molecular Sequence Data; D011550:Pseudomonas aeruginosa / Q000737:chemistry; D017386:Sequence Homology, Amino Acid; D012763:Shigella flexneri / Q000737:chemistry; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
D001426:Bacterial Proteins; C109404:CapB protein, Pseudomonas fragi; D002352:Carrier Proteins; D029968:Escherichia coli Proteins; D006360:Heat-Shock Proteins; C062260:cold shock protein CS7.4, Bacteria; C084501:hdeA protein, E coli; C084502:hdeB protein, E coli