Identification of glycoprotein components of alpha-agglutinin, a cell adhesion protein from Saccharomyces cerevisiae.
Several glycoproteins which inhibit the agglutinability of Saccharomyces cerevisiae mating type a cells were partially purified from extracts of mating type alpha cells. These proteins, called alpha-agglutinin, were labeled with 125I-Bolton-Hunter reagent. The labeled alpha-agglutinin showed specific binding to a cells. Such specific binding approached saturation with respect to agglutinin or cells and was inhibited in the presence of excess unlabeled alpha-agglutinin. Nonspecific binding was similar in a and alpha cells, was neither saturable nor competable, and was three- to fourfold less than the specific binding to a cells at maximum tested agglutinin concentrations. The major a-specific binding species had a low electrophoretic mobility in sodium dodecyl sulfate-polyacrylamide gels and had an apparent molecular weight of 155,000 by rate zonal centrifugation. Endo-N-acetylglucosaminidase H digestion of the purified glycoprotein complex converted the low-mobility material to four major and several minor bands which were resolved by polyacrylamide gel electrophoresis. All but two minor peptides bound specifically to a cells. Analyses of agglutinin from mnn mutants confirmed the deglycosylation results in suggesting that the N-linked carbohydrate portion of alpha-agglutinin was not necessary for activity.
D000371:Agglutination; D000373:Agglutinins / Q000302:isolation & purification* / Q000502:physiology; D005656:Fungal Proteins / Q000302:isolation & purification* / Q000502:physiology; D006023:Glycoproteins / Q000302:isolation & purification* / Q000502:physiology; D007457:Iodine Radioisotopes; D007700:Kinetics; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000502:physiology*; D013045:Species Specificity
D000373:Agglutinins; D005656:Fungal Proteins; D006023:Glycoproteins; D007457:Iodine Radioisotopes