title: Evidence for adhesin activity in the acid-stable moiety of the phosphomannoprotein cell wall complex of Candida albicans.
abstract: Previously, we showed that Candida albicans hydrophilic yeast cells adhere specifically to mouse splenic marginal-zone macrophages. The adhesins are part of the yeast cell wall phosphomannoprotein complex, and one adhesin site, which reacts with the monoclonal antibody 10G, was identified as a beta-1,2-linked tetramannose in the acid-labile portion of the complex. We report here that the acid-stable part of the complex, which has not been reported previously to have adhesin activity, is in large part responsible for yeast cell binding to the splenic marginal zone. The phosphomannoprotein complex, termed Fr.II, was isolated from C. albicans serotype B yeast cells by beta-mercaptoethanol extraction and concanavalin A-agarose affinity chromatography. Fr.II is devoid of the serotype A-specific antigen factor 6, which functions in yeast cell attachment to epithelial cells. The acid-stable part of Fr.II (i.e., Fr.IIS) was obtained by mild acid hydrolysis and size exclusion fractionation. Fr.IIS was further fractionated into four fractions, Fr.IIS1, Fr.IIS2, Fr.IIS3, and Fr.IIS4, by concanavalin A-agarose column chromatography and elution with a linear gradient of alpha-methyl-D-mannopyranoside. Adhesin activity of these fractions was determined by their ability to block yeast cell binding to the splenic marginal zone. Fr.IIS1 and Fr.IIS2 yielded more material and stronger adhesin activity than either Fr.IIS3 or Fr.IIS4. Only Fr.IIS1 did not react with antibodies (anti-factor 5 and monoclonal antibody 10G) specific for the acid-labile beta-1,2-linked oligosaccharides. Fr.IIS1-coated latex beads attached specifically to the marginal zone in a pattern identical to that of yeast cell binding. Furthermore, Fr.IIS1-latex bead attachment was inhibited by soluble Fr.II or Fr.IIS. Initial chemical analyses indicate that the adhesin site on Fr.IIS1 is a carbohydrate because adhesin activity was destroyed by periodate oxidation but not by proteinase K digestion.
authors.affiliation: Laboratory of Medical Mycology, Nagoya University School of Medicine, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Kanbe
authors.forename: J E
authors.initials: JE
authors.lastname: Cutler
mesh_terms: D000268:Adhesiveness; D002176:Candida albicans / Q000737:chemistry / Q000472:pathogenicity*; D002473:Cell Wall / Q000737:chemistry; D015151:Immunoblotting; D008562:Membrane Glycoproteins / Q000032:analysis*; D010750:Phosphoproteins / Q000032:analysis*
chemical_list: D008562:Membrane Glycoproteins; D010750:Phosphoproteins; C025363:mannoproteins