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: M200062-MCP200v1 1/12/967    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 Pitarch, A. Articles by Gil, C. Search for Related Content PubMed PubMed Citation Articles by Pitarch, A. Articles by Gil, C. Social Bookmarking                      What's this?

Molecular & Cellular Proteomics 1:967-982, 2002.
© 2002 by The American Society for Biochemistry and Molecular Biology, Inc.

---

Research

Sequential Fractionation and Two-dimensional Gel Analysis Unravels the Complexity of the Dimorphic Fungus Candida albicans Cell Wall Proteome^*

Aida Pitarch^,, Miguel Sánchez^¶, César Nombela^,|| and Concha Gil^,**

Departamento de Microbiología II, Facultad de Farmacia, Universidad Complutense de Madrid, 28040 Madrid, Spain
^¶ Departamento de Microbiología y Genética IMB-CSIC, Universidad de Salamanca, 37007 Salamanca, Spain

The cell wall proteins of Candida albicans play a key role in morphogenesis and pathogenesis and might be potential target sites for new specific antifungal drugs. However, these proteins are difficult to analyze because of their high heterogeneity, interconnections with wall polysaccharides (mannan, glucan, and chitin), low abundance, low solubility, and hydrophobic nature. Here we report a subproteomic approach for the study of the cell wall proteins (CWPs) from C. albicans yeast and hyphal forms. Most of the mannoproteins present in this compartment were extracted by cell wall fractionation according to the type of interactions that they establish with other structural components. CWPs were solubilized from isolated cell walls by hot SDS and dithiothreitol treatment followed by extraction either by mild alkali conditions or by enzymatic treatment with glucanases and chitinases. These highly enriched cell wall fractions were analyzed by two-dimensional PAGE, showing that a large number of proteins are involved in cell wall construction and that the wall remodeling that occurs during germ tube formation is related to changes in the composition of CWPs. We suggest that the CWP-chitin linkage is an important retention mechanism of CWPs in C. albicans mycelial forms. This article also highlights the usefulness of the combination of sequential fractionation and two-dimensional PAGE followed by Western blotting using specific antibodies against known CWPs in the characterization of incorporation mechanisms of such CWPs into the cell wall and of their interactions with other wall components. Mass spectrometry analyses have allowed the identification of several cell surface proteins classically associated with both the cell wall and other compartments. The physiological significance of the dual location of these moonlighting proteins is also discussed. This approach is therefore a powerful tool for obtaining a comprehensive and integrated view of the cell wall proteome.

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

This article has been cited by other articles:

				C. Fradin, M. C. Slomianny, C. Mille, A. Masset, R. Robert, B. Sendid, J. F. Ernst, J. C. Michalski, and D. Poulain {beta}-1,2 Oligomannose Adhesin Epitopes Are Widely Distributed over the Different Families of Candida albicans Cell Wall Mannoproteins and Are Associated through both N- and O-Glycosylation Processes Infect. Immun., October 1, 2008; 76(10): 4509 - 4517. [Abstract] [Full Text] [PDF]

				W. L. Chaffin Candida albicans Cell Wall Proteins Microbiol. Mol. Biol. Rev., September 1, 2008; 72(3): 495 - 544. [Abstract] [Full Text] [PDF]

				S. Poltermann, A. Kunert, M. von der Heide, R. Eck, A. Hartmann, and P. F. Zipfel Gpm1p Is a Factor H-, FHL-1-, and Plasminogen-binding Surface Protein of Candida albicans J. Biol. Chem., December 28, 2007; 282(52): 37537 - 37544. [Abstract] [Full Text] [PDF]

				J. Zhu, S. Alvarez, E. L. Marsh, M. E. LeNoble, I.-J. Cho, M. Sivaguru, S. Chen, H. T. Nguyen, Y. Wu, D. P. Schachtman, et al. Cell Wall Proteome in the Maize Primary Root Elongation Zone. II. Region-Specific Changes in Water Soluble and Lightly Ionically Bound Proteins under Water Deficit Plant Physiology, December 1, 2007; 145(4): 1533 - 1548. [Abstract] [Full Text] [PDF]

				C. Fradin, A. L. Mavor, G. Weindl, M. Schaller, K. Hanke, S. H. E. Kaufmann, H. Mollenkopf, and B. Hube The Early Transcriptional Response of Human Granulocytes to Infection with Candida albicans Is Not Essential for Killing but Reflects Cellular Communications Infect. Immun., March 1, 2007; 75(3): 1493 - 1501. [Abstract] [Full Text] [PDF]

				G. M. Olson, D. S. Fox, P. Wang, J. A. Alspaugh, and K. L. Buchanan Role of Protein O-Mannosyltransferase Pmt4 in the Morphogenesis and Virulence of Cryptococcus neoformans Eukaryot. Cell, February 1, 2007; 6(2): 222 - 234. [Abstract] [Full Text] [PDF]

				X. S. Li, J. N. Sun, K. Okamoto-Shibayama, and M. Edgerton Candida albicans Cell Wall Ssa Proteins Bind and Facilitate Import of Salivary Histatin 5 Required for Toxicity J. Biol. Chem., August 11, 2006; 281(32): 22453 - 22463. [Abstract] [Full Text] [PDF]

				P. K. Mukherjee, S. Mohamed, J. Chandra, D. Kuhn, S. Liu, O. S. Antar, R. Munyon, A. P. Mitchell, D. Andes, M. R. Chance, et al. Alcohol Dehydrogenase Restricts the Ability of the Pathogen Candida albicans To Form a Biofilm on Catheter Surfaces through an Ethanol-Based Mechanism Infect. Immun., July 1, 2006; 74(7): 3804 - 3816. [Abstract] [Full Text] [PDF]

				A. Pitarch, A. Jimenez, C. Nombela, and C. Gil Decoding Serological Response to Candida Cell Wall Immunome into Novel Diagnostic, Prognostic, and Therapeutic Candidates for Systemic Candidiasis by Proteomic and Bioinformatic Analyses Mol. Cell. Proteomics, January 1, 2006; 5(1): 79 - 96. [Abstract] [Full Text] [PDF]

				J. Zhu, S. Chen, S. Alvarez, V. S. Asirvatham, D. P. Schachtman, Y. Wu, and R. E. Sharp Cell Wall Proteome in the Maize Primary Root Elongation Zone. I. Extraction and Identification of Water-Soluble and Lightly Ionically Bound Proteins Plant Physiology, January 1, 2006; 140(1): 311 - 325. [Abstract] [Full Text] [PDF]

				N. Denikus, F. Orfaniotou, G. Wulf, P. F. Lehmann, M. Monod, and U. Reichard Fungal Antigens Expressed during Invasive Aspergillosis Infect. Immun., August 1, 2005; 73(8): 4704 - 4713. [Abstract] [Full Text] [PDF]

				A. Duran and C. Nombela Fungal cell wall biogenesis: building a dynamic interface with the environment Microbiology, October 1, 2004; 150(10): 3099 - 3103. [Full Text] [PDF]

				R. Martinez-Lopez, L. Monteoliva, R. Diez-Orejas, C. Nombela, and C. Gil The GPI-anchored protein CaEcm33p is required for cell wall integrity, morphogenesis and virulence in Candida albicans Microbiology, October 1, 2004; 150(10): 3341 - 3354. [Abstract] [Full Text] [PDF]

				P. W. J. de Groot, A. D. de Boer, J. Cunningham, H. L. Dekker, L. de Jong, K. J. Hellingwerf, C. de Koster, and F. M. Klis Proteomic Analysis of Candida albicans Cell Walls Reveals Covalently Bound Carbohydrate-Active Enzymes and Adhesins Eukaryot. Cell, August 1, 2004; 3(4): 955 - 965. [Abstract] [Full Text] [PDF]

				D. R. Singleton and K. C. Hazen Differential surface localization and temperature-dependent expression of the Candida albicans CSH1 protein Microbiology, February 1, 2004; 150(2): 285 - 292. [Abstract] [Full Text] [PDF]