Automated postprocessing of electrospray LC/MS data for profiling protein expression in bacteria

被引:34
作者
Williams, TL
Leopold, P
Musser, S
机构
[1] US FDA, Instrumentat & Biophys Branch, Ctr Food Safety & Appl Nutr, College Pk, MD 20740 USA
[2] BioAnalyte Inc, Portland, ME 04101 USA
关键词
D O I
10.1021/ac0258958
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
We describe an integrated approach for automating protein analysis of bacterial cell extracts. The method uses electrospray LC/MS to generate chromatographic profiles of proteins present in an extract, along with a software program that automates the data analysis. The software program, Retana, automates the sequential summing, centroiding, and deconvolution of multiply charged proteins present in consecutive scans of the LC/MS analysis. This procedure generates a concise, single spectrum of proteins present in the extract, along with their retention time and relative abundance. A comparison of the method with "whole cell" MALDI analysis demonstrates improved mass resolution and mass accuracy, along with the appearance of a greater number of proteins. Additionally, it is possible to compare protein expression among strains of bacteria by normalizing the relative abundance of similar proteins in each analysis.
引用
收藏
页码:5807 / 5813
页数:7
相关论文
共 45 条
[1]   Observation of Escherichia coli ribosomal proteins and their posttranslational modifications by mass spectrometry [J].
Arnold, RJ ;
Reilly, JP .
ANALYTICAL BIOCHEMISTRY, 1999, 269 (01) :105-112
[2]  
Birmingham J, 1999, RAPID COMMUN MASS SP, V13, P604, DOI 10.1002/(SICI)1097-0231(19990415)13:7<604::AID-RCM529>3.0.CO
[3]  
2-M
[4]  
Chen YJ, 1998, RAPID COMMUN MASS SP, V12, P1994, DOI 10.1002/(SICI)1097-0231(19981230)12:24<1994::AID-RCM423>3.0.CO
[5]  
2-2
[6]  
Chong BE, 1999, RAPID COMMUN MASS SP, V13, P1808, DOI 10.1002/(SICI)1097-0231(19990930)13:18<1808::AID-RCM720>3.0.CO
[7]  
2-U
[8]   Chromatofocusing nonporous reversed-phase high-performance liquid chromatography/electrospray ionization time-of-flight mass spectrometry of proteins from human breast cancer whole cell lysates: a novel two-dimensional liquid chromatography/mass spectrometry method [J].
Chong, BE ;
Yan, F ;
Lubman, DM ;
Miller, FR .
RAPID COMMUNICATIONS IN MASS SPECTROMETRY, 2001, 15 (04) :291-296
[9]   Differential screening and mass mapping of proteins from premalignant and cancer cell lines using nonporous reversed-phase HPLC coupled with mass spectrometric analysis [J].
Chong, BE ;
Hamler, RL ;
Lubman, DM ;
Ethier, SP ;
Rosenspire, AJ ;
Miller, FR .
ANALYTICAL CHEMISTRY, 2001, 73 (06) :1219-1227
[10]   Use of non-porous reversed-phase high-performance liquid chromatography for protein profiling and isolation of proteins induced by temperature variations for Siberian permafrost bacteria with identification by matrix-assisted laser desorption lionization time-of-flight mass spectrometry and capillary electrophoresis-electrospray ionization mass spectrometry [J].
Chong, BE ;
Kim, J ;
Lubman, DM ;
Tiedje, JM ;
Kathariou, S .
JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES, 2000, 748 (01) :167-177