Saccharomyces cerevisiae a- and alpha-agglutinin: characterization of their molecular interaction.
An O-glycosylated protein of approximately 18 kDa responsible for mating type specific agglutination has been isolated from Saccharomyces cerevisiae a cells, purified to homogeneity and via peptide sequences the gene was cloned by PCR. An open reading frame codes for a protein of 69 amino acids. A minimum of five serine and five threonine residues of the mature protein are glycosylated. alpha-Agglutinin is a highly N-glycosylated protein of approximately 250 kDa. Both purified agglutinins form a specific 1:1 complex in vitro. Pretreatment of alpha-agglutinin, but not of alpha-agglutinin, with diethylpyrocarbonate (DEPC) prevents formation of the complex; treatment of alpha-agglutinin in the presence of alpha-agglutinin protects the former from DEPC inactivation. By carboxy terminal shortening of the alpha-agglutinin gene and by replacing three of its eight histidyl residues by arginine, the active region of alpha-agglutinin for interaction with alpha-agglutinin has been defined. Neither the N- nor the O-linked saccharides of the two agglutinins seem to be essential for their interaction.
Cappellaro
C
C
Lehrstuhl für Zellbiologie und Pflanzenphysiologie, Universität Regensburg, FRG.
Hauser
K
K
Mrśa
V
V
Watzele
M
M
Watzele
G
G
Gruber
C
C
Tanner
W
W
D001483:Base Sequence; D015152:Blotting, Northern; D003001:Cloning, Molecular; D004047:Diethyl Pyrocarbonate / Q000737:chemistry; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism*; D006031:Glycosylation; D000072235:Mating Factor; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D016366:Open Reading Frames; D010449:Peptide Mapping; D010455:Peptides / Q000235:genetics / Q000378:metabolism*; D010675:Pheromones / Q000378:metabolism; D016133:Polymerase Chain Reaction; D012331:RNA, Fungal / Q000032:analysis; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins; D014357:Trypsin
D005656:Fungal Proteins; D010455:Peptides; D010675:Pheromones; D012331:RNA, Fungal; D029701:Saccharomyces cerevisiae Proteins; C069939:a-agglutinin protein, S cerevisiae; D000072235:Mating Factor; D014357:Trypsin; D004047:Diethyl Pyrocarbonate