Burt E T, O'Connor C, Larsen B
Department of Biochemistry, Des Moines University-Osteopathic Medical Center, IA 50312, USA.
Mycopathologia. 1999;147(1):13-20. doi: 10.1023/a:1007036518330.
It was previously shown that the presence of estrogen enhances survival of Candida albicans under heat and oxidative stresses. A 92-kDa protein is inducible by heat shock and estrogen in C. albicans. Previous studies have described this protein as hsp90 because of its molecular size and heat inducibility as seen on electrophoretic gels and Western blots. In this study, ion exchange, hydroxyapatite and size exclusion chromatography were used to isolate a 92-kDa-protein band. The N-terminal sequence of isolated protein blotted onto a PVDF membrane was determined to be V-Q-S-?-V-L-G-F-P-R. This sequence is homologous to the N-terminal sequence of the MET6 gene product, cobalamin-independent methionine synthase, from Saccharomyces cerevisiae. The results of this study suggest that a cobalamin-independent methionine synthase homolog is inducible by heat and estrogen in C. albicans. This study also suggests that Candida hsp90 is more likely to exist as an 82-kDa protein as predicted by a previously described cDNA and not as a 92-kDa protein as reported in the literature.
先前的研究表明,雌激素的存在可提高白色念珠菌在热应激和氧化应激下的存活率。在白色念珠菌中,一种92 kDa的蛋白质可被热休克和雌激素诱导产生。由于其分子大小以及在电泳凝胶和蛋白质免疫印迹上呈现的热诱导性,先前的研究将这种蛋白质描述为热休克蛋白90(hsp90)。在本研究中,采用离子交换、羟基磷灰石和尺寸排阻色谱法分离出一条92 kDa的蛋白带。经测定,转移至聚偏二氟乙烯(PVDF)膜上的分离蛋白的N端序列为V-Q-S-?-V-L-G-F-P-R。该序列与酿酒酵母中钴胺素非依赖性甲硫氨酸合酶MET6基因产物的N端序列同源。本研究结果表明,白色念珠菌中一种钴胺素非依赖性甲硫氨酸合酶同源物可被热和雌激素诱导产生。本研究还表明,白色念珠菌热休克蛋白90更可能如先前所述的cDNA所预测的那样,以82 kDa的蛋白质形式存在,而不是如文献报道的92 kDa蛋白质。