Eroles Pilar, Sentandreu Maria, Elorza M Victoria, Sentandreu Rafael
Secci Departamental de Microbiologa, Facultat de Farmcia, Universitat de Valncia, Avgda. V. Andrs Estelles s/n, 46100 Burjassot, Valncia, Spain.
Microbiology (Reading). 1997 Feb;143 ( Pt 2):313-320. doi: 10.1099/00221287-143-2-313.
Screening cDNA libraries with polyclonal antibody preparations against Candida albicans yeast or mycelial cell walls resulted in isolation of several positive clones. Some of them encoded enolase; others encoded a protein of the 70 kDa heat-shock protein family (Hsp70p), etc. The presence of these cytosolic proteins in the cell wall of actively growing C. albicans was discovered by analytical (SDS-PAGE and Western blot) and cytological (indirect immunofluorescence) experiments. Supplementation of cell cultures with papulacandin B, an antibiotic that inhibits formation of the beta-glucan skeleton, resulted in the release of enolase to the supernatant fluids; this release was prevented when 0.6 M KCl was present as an osmotic stabilizer. The cell wall of C. albicans incorporated exogenously added proteins (enolase and Escherichia coli and C., albicans cytosolic proteins). The presence in the C. albicans cell wall of enolase, Hsp70p, and probably other intracellular proteins that are highly immunogenic might help the fungal cells to evade the host defences, and consequently could represent a survival mechanism for C. albicans 'in vivo'.
用针对白色念珠菌酵母或菌丝细胞壁的多克隆抗体制剂筛选cDNA文库,结果分离出几个阳性克隆。其中一些编码烯醇化酶;其他的编码70 kDa热休克蛋白家族(Hsp70p)的一种蛋白质等。通过分析实验(SDS - PAGE和蛋白质印迹法)和细胞学实验(间接免疫荧光法)发现,这些胞质蛋白存在于活跃生长的白色念珠菌的细胞壁中。用抑制β - 葡聚糖骨架形成的抗生素papulacandin B补充细胞培养物,导致烯醇化酶释放到上清液中;当存在0.6 M KCl作为渗透稳定剂时,这种释放被阻止。白色念珠菌的细胞壁整合了外源添加的蛋白质(烯醇化酶、大肠杆菌和白色念珠菌的胞质蛋白)。烯醇化酶、Hsp70p以及可能其他具有高度免疫原性的细胞内蛋白质存在于白色念珠菌的细胞壁中,这可能有助于真菌细胞逃避宿主防御,因此可能代表白色念珠菌在“体内”的一种生存机制。