HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) All Versions of this Article: M300074-MCP200v1 3/1/24    most recent Submit a response Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Glossary Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Schmidt, F. Articles by Jungblut, P. R. Search for Related Content PubMed PubMed Citation Articles by Schmidt, F. Articles by Jungblut, P. R. Social Bookmarking                      What's this?

Molecular & Cellular Proteomics 3:24-42, 2004.
© 2004 by The American Society for Biochemistry and Molecular Biology, Inc.

---

Research

Complementary Analysis of the Mycobacterium tuberculosis Proteome by Two-dimensional Electrophoresis and Isotope-coded Affinity Tag Technology ^*

Frank Schmidt, Samuel Donahoe, Kristine Hagens^¶, Jens Mattow^¶, Ulrich E. Schaible^¶, Stefan H. E. Kaufmann^¶, Ruedi Aebersold and Peter R. Jungblut^,||

From the Core Facility Protein Analysis and ^¶ Department of Immunology, Max Planck Institute for Infection Biology, D-10117 Berlin, Germany and the Institute for Systems Biology, Seattle, Washington 98103-8904

Classical proteomics combined two-dimensional gel electrophoresis (2-DE) for the separation and quantification of proteins in a complex mixture with mass spectrometric identification of selected proteins. More recently, the combination of liquid chromatography (LC), stable isotope tagging, and tandem mass spectrometry (MS/MS) has emerged as an alternative quantitative proteomics technology. We have analyzed the proteome of Mycobacterium tuberculosis, a major human pathogen comprising about 4,000 genes, by (i) 2-DE and mass spectrometry (MS) and by (ii) the isotope-coded affinity tag (ICAT) reagent method and MS/MS. The data obtained by either technology were compared with respect to their selectivity for certain protein types and classes and with respect to the accuracy of quantification. Initial datasets of 60,000 peptide MS/MS spectra and 1,800 spots for the ICAT-LC/MS and 2-DE/MS methods, respectively, were reduced to 280 and 108 conclusively identified and quantified proteins, respectively. ICAT-LC/MS showed a clear bias for high M_r proteins and was complemented by the 2-DE/MS method, which showed a preference for low M_r proteins and also identified cysteine-free proteins that were transparent to the ICAT-LC/MS method. Relative quantification between two strains of the M. tuberculosis complex also revealed that the two technologies provide complementary quantitative information; whereas the ICAT-LC/MS method quantifies the sum of the protein species of one gene product, the 2-DE/MS method quantifies at the level of resolved protein species, including post-translationally modified and processed polypeptides. Our data indicate that different proteomic technologies applied to the same sample provide complementary types of information that contribute to a more complete understanding of the biological system studied.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				C. S. Andersson and M. Hogbom A Mycobacterium tuberculosis ligand-binding Mn/Fe protein reveals a new cofactor in a remodeled R2-protein scaffold PNAS, April 7, 2009; 106(14): 5633 - 5638. [Abstract] [Full Text] [PDF]

				J. A. McDonough, J. R. McCann, E. M. Tekippe, J. S. Silverman, N. W. Rigel, and M. Braunstein Identification of Functional Tat Signal Sequences in Mycobacterium tuberculosis Proteins J. Bacteriol., October 1, 2008; 190(19): 6428 - 6438. [Abstract] [Full Text] [PDF]

				S. C. G. Rison, J. Mattow, P. R. Jungblut, and N. G. Stoker Experimental determination of translational starts using peptide mass mapping and tandem mass spectrometry within the proteome of Mycobacterium tuberculosis Microbiology, February 1, 2007; 153(2): 521 - 528. [Abstract] [Full Text] [PDF]

				J. Yuan, L. Zhu, X. Liu, T. Li, Y. Zhang, T. Ying, B. Wang, J. Wang, H. Dong, E. Feng, et al. A Proteome Reference Map and Proteomic Analysis of Bifidobacterium longum NCC2705 Mol. Cell. Proteomics, June 1, 2006; 5(6): 1105 - 1118. [Abstract] [Full Text] [PDF]

				B. Canas, D. Lopez-Ferrer, A. Ramos-Fernandez, E. Camafeita, and E. Calvo Mass spectrometry technologies for proteomics Briefings in Functional Genomics, February 1, 2006; 4(4): 295 - 320. [Abstract] [Full Text] [PDF]

				K. K. Singh, Y. Dong, S. A. Patibandla, D. N. McMurray, V. K. Arora, and S. Laal Immunogenicity of the Mycobacterium tuberculosis PPE55 (Rv3347c) Protein during Incipient and Clinical Tuberculosis Infect. Immun., August 1, 2005; 73(8): 5004 - 5014. [Abstract] [Full Text] [PDF]

				R. Wang, J. T. Prince, and E. M. Marcotte Mass spectrometry of the M. smegmatis proteome: Protein expression levels correlate with function, operons, and codon bias Genome Res., August 1, 2005; 15(8): 1118 - 1126. [Abstract] [Full Text] [PDF]

				S. Sinha, K. Kosalai, S. Arora, A. Namane, P. Sharma, A. N. Gaikwad, P. Brodin, and S. T. Cole Immunogenic membrane-associated proteins of Mycobacterium tuberculosis revealed by proteomics Microbiology, July 1, 2005; 151(7): 2411 - 2419. [Abstract] [Full Text] [PDF]

				X.-S. Jiang, L.-Y. Tang, J. Dai, H. Zhou, S.-J. Li, Q.-C. Xia, J.-R. Wu, and R. Zeng Quantitative Analysis of Severe Acute Respiratory Syndrome (SARS)-associated Coronavirus-infected Cells Using Proteomic Approaches: Implications for Cellular Responses to Virus Infection Mol. Cell. Proteomics, July 1, 2005; 4(7): 902 - 913. [Abstract] [Full Text] [PDF]

				X.-S. Jiang, J. Dai, Q.-H. Sheng, L. Zhang, Q.-C. Xia, J.-R. Wu, and R. Zeng A Comparative Proteomic Strategy for Subcellular Proteome Research: Icat Approach Coupled with Bioinformatics Prediction to Ascertain Rat Liver Mitochondrial Proteins and Indication of Mitochondrial Localization for Catalase Mol. Cell. Proteomics, January 1, 2005; 4(1): 12 - 34. [Abstract] [Full Text] [PDF]

				J. Wissing, K. Godl, D. Brehmer, S. Blencke, M. Weber, P. Habenberger, M. Stein-Gerlach, A. Missio, M. Cotten, S. Muller, et al. Chemical Proteomic Analysis Reveals Alternative Modes of Action for Pyrido[2,3-d]pyrimidine Kinase Inhibitors Mol. Cell. Proteomics, December 1, 2004; 3(12): 1181 - 1193. [Abstract] [Full Text] [PDF]

				C. Li, Y. Hong, Y.-X. Tan, H. Zhou, J.-H. Ai, S.-J. Li, L. Zhang, Q.-C. Xia, J.-R. Wu, H.-Y. Wang, et al. Accurate Qualitative and Quantitative Proteomic Analysis of Clinical Hepatocellular Carcinoma Using Laser Capture Microdissection Coupled with Isotope-coded Affinity Tag and Two-dimensional Liquid Chromatography Mass Spectrometry Mol. Cell. Proteomics, April 1, 2004; 3(4): 399 - 409. [Abstract] [Full Text] [PDF]