MALDI mass spectrometry analysis of high molecular weight proteins from whole bacterial cells: pretreatment of samples with surfactants.
The use of surfactants as additives in conjunction with on-probe whole cell bacterial protein analysis employing MALDI-TOF-MS is described. Nonionic and zwitterionic surfactants were used to enhance the detection of high molecular weight proteins. Three nonionic, N-octyl-B-D-glactopyranoside, N-decyl-B-D-maltopyranoside, and N-dodecyl-B-D-maltoside, and two zwitterionic surfactants, N,N-dimethyldodecylamine-N-oxide and zwittergent 3-12 were evaluated with five different MALDI matrix systems. New peaks in the mass range of 2 to 80 kDa were produced with all of the various combinations of matrix and surfactant from both whole cell gram-positive and gram-negative bacteria. Ferulic acid used in conjunction with a 1.0 mM solution of N-octyl-B-D-glactopyranoside produced the highest quality spectra with high signal to noise ratios and peaks up to 140 kDa.
Meetani
Mohammed A
MA
Department of Chemistry and Geochemistry, Colorado School of Mines, 14th and Illinois St., 80401, Golden, CO, USA.
Voorhees
Kent J
KJ
Department of Chemistry and Geochemistry, Colorado School of Mines, 14th and Illinois St., 80401, Golden, CO, USA. kvoorhee@mines.edu.
D001419:Bacteria / Q000737:chemistry* / Q000378:metabolism*; D001426:Bacterial Proteins / Q000737:chemistry*; D018929:Cell Culture Techniques / Q000379:methods*; D008970:Molecular Weight; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013048:Specimen Handling / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D013501:Surface-Active Agents / Q000737:chemistry*
D001426:Bacterial Proteins; D013501:Surface-Active Agents