Burghoorn Henriette P, Soteropoulos Patricia, Paderu Padmaja, Kashiwazaki Ryota, Perlin David S
Public Health Research Institute, New York, New York 10016, USA.
Antimicrob Agents Chemother. 2002 Mar;46(3):615-24. doi: 10.1128/AAC.46.3.615-624.2002.
The gene encoding the plasma membrane proton pump (H+ -ATPase) of Aspergillus fumigatus, PMA1, was characterized from A. fumigatus strain NIH 5233 and clinical isolate H11-20. An open reading frame of 3,109 nucleotides with two introns near the N terminus predicts a protein consisting of 989 amino acids with a molecular mass of approximately 108 kDa. The predicted A. fumigatus enzyme is 89 and 51% identical to H+ - ATPases of Aspergillus nidulans and Saccharomyces cerevisiae, respectively. The A. fumigatus PMA1 is a typical member of the P-type ATPase family that contains 10 predicted transmembrane segments and conserved sequence motifs TGES, CSDKTGT, MLTGD, and GDGVN within the catalytic region. The enzyme represents 2% of the total plasma membrane protein, and it is characteristically inhibited by orthovanadate, with a 50% inhibitory concentration of approximately 1.8 microM. H+ -ATPases from Aspergillus spp. contain a highly acidic insertion region of 60 amino acids between transmembrane segments 2 and 3, which was confirmed for the membrane-assembled enzyme with a peptide-derived antibody. An increasing A. fumigatus PMA1 copy number confers enhanced growth in low-pH medium, consistent with its role as a proton pump. These results provide support for the development of the A. fumigatus H+ -ATPase as a potential drug discovery target.
对烟曲霉的质膜质子泵（H⁺-ATP 酶）编码基因 PMA1，从烟曲霉 NIH 5233 菌株和临床分离株 H11-20 进行了特性分析。一个 3109 个核苷酸的开放阅读框在 N 端附近有两个内含子，预测编码一个由 989 个氨基酸组成、分子量约为 108 kDa 的蛋白质。预测的烟曲霉酶与构巢曲霉和酿酒酵母的 H⁺-ATP 酶分别有 89%和 51%的同一性。烟曲霉 PMA1 是 P 型 ATP 酶家族的典型成员，在催化区域包含 10 个预测的跨膜片段以及保守序列基序 TGES、CSDKTGT、MLTGD 和 GDGVN。该酶占质膜总蛋白的 2%，其特征性地被原钒酸盐抑制，50%抑制浓度约为 1.8 μM。曲霉属的 H⁺-ATP 酶在跨膜片段 2 和 3 之间含有一个 60 个氨基酸的高度酸性插入区域，这通过肽衍生抗体对膜组装酶得到了证实。烟曲霉 PMA1 拷贝数增加赋予其在低 pH 培养基中增强的生长能力，与其作为质子泵的作用一致。这些结果为将烟曲霉 H⁺-ATP 酶开发为潜在的药物发现靶点提供了支持。
