The beta-aspartyl phosphate intermediate in a Leishmania donovani promastigote plasma membrane P-type ATPase.
The beta-aspartyl phosphate intermediate in a Leishmania donovani promastigote plasma membrane P-type ATPase.
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杜氏利什曼原虫前鞭毛体质膜 P 型 ATP 酶中的 β-天冬氨酰磷酸中间体。
DOI：
10.1016/0005-2736(94)90012-4
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发表时间：
1994
期刊：
影响因子：
--
通讯作者：
Mukkada,AJ
中科院分区：
文献类型：
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作者：
Anderson,SA;Jiang,S;Mukkada,AJ
进一步鉴定了杜氏利什曼原虫质膜P型ATPase的磷酸化中间体。磷酸化中间体的形成对几种ATPase抑制剂敏感，包括钒酸盐、二环己基碳二亚胺(DCCD)、N-乙基马来酰亚胺(NEM)和异硫氰酸荧光素(FITC)。这些抑制物影响纯化的免疫沉淀蛋白以及总的质膜部分。线粒体ATPase抑制剂寡霉素和Na+/K+-ATPase抑制剂哇巴因对磷酸化中间体的形成没有影响。ATPase磷酸蛋白是酸性稳定的，在碱性pH下脱磷，表明酰基磷酸化学键的存在。用硼氢化钠还原磷酸化氨基酸的分析表明，残基是天冬氨酸，证实了β-天冬氨酸磷酸酯中间体的形成。这些数据表明，杜氏乳杆菌质膜上存在一个105 kDa的P型ATPase，它与高等真核生物中的其他P型酶在力学上是相似的。
The phosphorylated intermediate of a plasma membrane P-type ATPase in Leishmania donovani has been further characterized. The formation of the phosphorylated intermediate is sensitive to several ATPase inhibitors including vanadate, dicyclohexyl carbodiimide (DCCD), N-ethylmaleimide (NEM), and fluorescein isothiocyanate (FITC). These inhibitors affect purified immunoprecipitated protein as well as total plasma membrane fractions. Oligomycin, an inhibitor of mitochondrial ATPases, and ouabain, an inhibitor of Na+/K+-ATPases, had no effect on the formation of the phosphorylated intermediate. The ATPase phosphoprotein was acid stable and dephosphorylated at alkaline pH, indicating the presence of the acyl phosphate chemical linkage. Analysis of the phosphorylated amino acid by reduction with sodium boro [3 H] hydride identified the residue as aspartate, confirming the formation of a β-aspartyl phosphate intermediate. These data indicate the presence of a 105 kDa P-type ATPase on L. donovani plasma membrane that is mechanistically similar to other P-type enzymes of higher eukaryotes.