Proteomic analysis of novel marine bacteria using MALDI and ESI mass spectrometry.
The objective of this study was to develop a mass spectrometric protocol to search for proteins related to phototrophy in marine bacteria. The genes that produce proteins involved in conversion of light into energy have been detected by cloning-sequencing from some of these bacteria, but it was previously unknown if these proteins were actually expressed. Attaining this study's goal was complicated by the fact that the samples consisted of miniscule cell pellets, which yielded small amounts of very complex mixtures of proteins. Sample preparation and analysis were tailored to optimize the probability of detecting the proteins of interest. It has been reported that using both matrix-assisted laser desorption/ionization (MALDI) and electrospray ionization (ESI) to analyze a mixture of peptides leads to the identification of more peptides that either technique alone. In order to exploit this complementarity between ESI and MALDI for proteomic analysis, samples were analyzed using both ionization techniques. With correct choices in sample preparation and ionization process, biologically relevant proteins can be identified out of small samples containing whole proteomes.
Stapels
Martha D
MD
Department of Chemistry, 153 Gilbert Hall, Oregon State University, Corvallis, OR 97331-4003, USA.
Cho
Jang-Cheon
JC
Giovannoni
Stephen J
SJ
Barofsky
Douglas F
DF
D020561:Alphaproteobacteria / Q000378:metabolism*; D000595:Amino Acid Sequence; D020563:Gammaproteobacteria / Q000378:metabolism*; D008969:Molecular Sequence Data; D020543:Proteome / Q000378:metabolism*; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
D020543:Proteome