Characterization of two Candida albicans surface mannoprotein adhesins that bind immobilized saliva components.
Salivary components, including the basic proline-rich proteins (bPRP), act as receptors for the adherence of Candida albicans, and this interaction may be important for oral colonization and the development of mucosal Candida infections. Treatment of C. albicans cells with agents affecting either proteinacious or carbohydrate surface macromolecules reduced their adherence in in vitro assays of C. albicans adherence to saliva-coated hydroxyapatite beads and to membrane-immobilized salivary bPRP. In order to identify C. albicans adhesins that bind saliva receptors, yeast cell surface material was extracted by mild glucanase treatment, and was shown to competitively inhibit ( > 50%) the adherence of C. albicans yeast cells in both assays. Concanavalin A sepharose affinity chromatography was used to partially purify glycosylated components of the extract, and two polypeptides (97.4 and 35 kDa) were further purified by preparative SDS PAGE separation and electro-elution. The 97.4 and 35 kDa polypetides each possessed greater adherence-inhibitory specific activity (> 100-fold and > 30-fold respectively) than the original glucanase extract from C. albicans yeast cells. The 35 kDa putative surface protein was identified by N-terminal sequencing and immunoblotting, as the 1,3-beta glucosyltransferase, Bgl2p.
Jeng
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HW
Department of Oral Sciences, School of Dentistry, University of Otago, Dunedin, New Zealand.
Holmes
A R
AR
Cannon
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RD
D000595:Amino Acid Sequence; D002176:Candida albicans / Q000737:chemistry* / Q000378:metabolism; D015815:Cell Adhesion Molecules; D002846:Chromatography, Affinity; D003910:Dextranase / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D006031:Glycosylation; D008562:Membrane Glycoproteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D008565:Membrane Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010455:Peptides / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D055232:Proline-Rich Protein Domains; D011485:Protein Binding; D012471:Salivary Proteins and Peptides / Q000378:metabolism*
D015815:Cell Adhesion Molecules; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D010455:Peptides; D012471:Salivary Proteins and Peptides; C025363:mannoproteins; D003910:Dextranase