Tosh F D, Douglas L J
Department of Microbiology, University of Glasgow, United Kingdom.
Infect Immun. 1992 Nov;60(11):4734-9. doi: 10.1128/iai.60.11.4734-4739.1992.
Candida albicans GDH 2346 produces extracellular polymeric material (EP) which contains a mannoprotein adhesin with a lectin-like affinity for fucose-containing glycosides. EP isolated from culture supernatants of this strain was used as starting material for purification of the adhesin. The purification protocol involved a stepwise treatment of EP with N-glycanase, papain, and dilute alkali to cleave the protein and carbohydrate portions of the mannoprotein molecule. Fucoside-binding protein fragments were then recovered by affinity adsorption with the trisaccharide determinant of the H (type 2) blood group antigen which terminates in a residue of L-fucose. The purified adhesin was devoid of carbohydrate and inhibited yeast adhesion to buccal epithelial cells 221 times more efficiently, on a protein weight basis, than did EP. Adhesion inhibition reached a maximum of 78 to 80% at an adhesin concentration of 10 micrograms ml-1. Our results indicate that this protein is the major adhesin of yeast-phase cells of C. albicans GDH 2346 but that one or more secondary adhesion mechanisms may operate.
白色念珠菌GDH 2346产生细胞外聚合物(EP),其中含有一种甘露糖蛋白黏附素,对含岩藻糖的糖苷具有类似凝集素的亲和力。从该菌株培养上清液中分离出的EP用作纯化黏附素的起始材料。纯化方案包括用N-聚糖酶、木瓜蛋白酶和稀碱对EP进行逐步处理,以切割甘露糖蛋白分子的蛋白质和碳水化合物部分。然后通过与H(2型)血型抗原的三糖决定簇进行亲和吸附来回收岩藻糖苷结合蛋白片段,该决定簇以L-岩藻糖残基结尾。纯化后的黏附素不含碳水化合物,以蛋白质重量计,其抑制酵母与颊上皮细胞黏附的效率比EP高221倍。在黏附素浓度为10微克/毫升时,黏附抑制率最高可达78%至80%。我们的结果表明,这种蛋白质是白色念珠菌GDH 2346酵母相细胞的主要黏附素,但可能存在一种或多种次要的黏附机制。