Isolation and biochemical characterization of the iC3b receptor of Candida albicans.
In an effort to identify the protein structure on Candida albicans, pseudohyphal forms which had been shown earlier to bind human iC3b, a protein of about 42 kDa (p42), were obtained from lysates of pseudohyphal forms by absorption with C3(H2O)-Sepharose. An antiserum raised in rabbits against this protein effectively inhibited adherence of sheep erythrocytes carrying iC3b (EAC3bi) to pseudohyphal forms. p42 cross-reacted with OKM-1, a monoclonal antibody directed against the human complement receptor type 3 (CR3, CD11b). This protein, p42, was designated p42-CR3. The antiserum against p42-CR3 was used for further purification of lysates by affinity chromatography. Three proteins of 66, 55, and 42 kDa were isolated. All were recognized by OKM-1 in immunoblots (p66-, p55-, and p42-CR3). The different proteins were separated and treated with neuraminidase and endoglycosidase F. Almost complete deglycosylation of the p66-CR3 protein was obtained after treatment with neuraminidase, indicating a high degree of glycosylation. Neuraminidase also had an effect on p55-CR3, but not on p42-CR3. Endoglycosidase F did not alter any of the three proteins. In ligand blots, p42-CR3 bound C3(H2O), C3b, and iC3b but not C3d; p55-CR3 clearly reacted with C3(H2O) and weakly reacted with C3b and iC3b. p66-CR3 never showed reactivity. It is suggested that p55 and p66 represent glycosylated forms of p42-CR3. Although C. albicans CR3 and human CR3 cross-react and bind identical ligands, the two receptors differ in structure.
Alaei
S
S
Institut für Hygiene, University of Innsbruck, Austria.
Larcher
C
C
Ebenbichler
C
C
Prodinger
W M
WM
Janatova
J
J
Dierich
M P
MP
D002176:Candida albicans / Q000737:chemistry / Q000276:immunology*; D002846:Chromatography, Affinity; D003176:Complement C3 / Q000378:metabolism; D005656:Fungal Proteins / Q000276:immunology / Q000302:isolation & purification; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase / Q000494:pharmacology; D009439:Neuraminidase / Q000494:pharmacology; D017463:Receptors, Complement 3b / Q000737:chemistry / Q000302:isolation & purification*
D003176:Complement C3; D005656:Fungal Proteins; D017463:Receptors, Complement 3b; D009439:Neuraminidase; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase