Glucan synthase complex of Aspergillus fumigatus
Glucan synthase complex of Aspergillus fumigatus
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DOI：
10.1128/jb.183.7.2273-2279.2001
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发表时间：
2001-04-01
影响因子：
3.2
通讯作者：
Latgé, JP
中科院分区：
文献类型：
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作者：
Beauvais, A;Bruneau, JM;Latgé, JP
对人体致病霉菌烟曲霉的葡聚糖合成酶复合体进行了研究。克隆了烟曲霉菌催化亚基Fks1p和4个Rho蛋白的编码基因，并对其进行了序列分析。序列分析表明，AfFks1p是一种跨膜蛋白，与酵母和尼杜拉曲霉中的其他Fksp蛋白非常相似。AFFks1p保守的内部亲水结构域在大肠杆菌中的异源表达抗Fks1p抗体标记胚管顶端，苯胺蓝荧光色素标记β(1-3)葡聚糖，表明AfFks1p与新合成的β(1-3)葡聚糖共定位。首次在丝状真菌中研究了与酿酒酵母RHO1同源性最高的基因AfRHO1。AfRho蛋白具有与酵母Rho蛋白相同的GTP结合和水解共识序列，并且在AfRho1p和AfRho3p中有一个略有修改的杰尔尼化位点。产物包埋法对葡聚糖合成酶复合体进行纯化，得到了四种蛋白：Fks1p、Rho1p，一个与膜HF-ATPase同源的100 kDa蛋白，以及一个由抗β(13)葡聚糖抗体标记的、与ABC细菌β(1-2)葡聚糖转运蛋白同源的160 kDa蛋白。
The glucan synthase complex of the human pathogenic mold Aspergillus fumigatus has been investigated. The genes encoding the putative catalytic subunit Fks1p and four Rho proteins of A. fumigatus were cloned and sequenced. Sequence analysis showed that AfFks1p was a transmembrane protein very similar to other Fksp proteins in yeasts and in Aspergillus nidulans. Heterologous expression of the conserved internal hydrophilic domain of AfFks1p aas achieved in Escherichia coli. Anti-Fks1p antibodies labeled the apex of the germ tube, as did aniline blue fluorochrome, which was specific for beta (1-3) glucans, showing that AfFks1p colocalized with the newly synthesized beta (1-3) glucans. AfRHO1, the most homologous gene to RHO1 of Saccharomyces cerevisiae, was studied for the first time in a filamentous fungus. AfRho proteins have GTP binding and hydrolysis consensus sequences identical to those of yeast Rho proteins and have a slightly modified geranylation site in AfRho1p and AfRho3p. Purification of the glucan synthase complex by product entrapment led to the enrichment of four proteins: Fks1p, Rho1p, a 100-kDa protein homologous to a membrane Hf-ATPase, and a 160-kDa protein which was labeled by an anti-beta (13) glucan antibody and was homologous to ABC bacterial beta (1-2) glucan transporters.