Cappellaro C, Hauser K, Mrśa V, Watzele M, Watzele G, Gruber C, Tanner W
Lehrstuhl für Zellbiologie und Pflanzenphysiologie, Universität Regensburg, FRG.
EMBO J. 1991 Dec;10(13):4081-8. doi: 10.1002/j.1460-2075.1991.tb04984.x.
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.
一种负责交配型特异性凝集的约18 kDa的O-糖基化蛋白已从酿酒酵母a细胞中分离出来，纯化至同质，并通过肽序列利用聚合酶链反应（PCR）克隆了该基因。一个开放阅读框编码一种69个氨基酸的蛋白质。成熟蛋白至少有五个丝氨酸和五个苏氨酸残基被糖基化。α-凝集素是一种高度N-糖基化的约250 kDa的蛋白质。两种纯化的凝集素在体外形成特定的1:1复合物。用焦碳酸二乙酯（DEPC）预处理α-凝集素（而非α-凝集素）可阻止复合物的形成；在α-凝集素存在的情况下处理α-凝集素可保护前者免受DEPC失活。通过α-凝集素基因的羧基末端缩短以及将其八个组氨酸残基中的三个替换为精氨酸，已确定了α-凝集素与α-凝集素相互作用的活性区域。两种凝集素的N-连接和O-连接糖似乎对它们的相互作用都不是必需的。
