Interaction between the N-terminal Domain of Gastric H,K-ATPase and the Spectrin Binding Domain of Ankyrin III*

Interaction between the N-terminal Domain of Gastric H,K-ATPase and the Spectrin Binding Domain of Ankyrin III*
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胃 H,K-ATP 酶 N 末端结构域与锚蛋白 III 血影蛋白结合结构域之间的相互作用*

DOI:
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发表时间:
2001
影响因子:
4.8
通讯作者:
Annick Thomas
Annick Thomas
中科院分区:
生物学2区
文献类型:
--
作者:
F. Festy;J. Robert;R. Brasseur;Annick Thomas

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我们通过双杂交技术筛选了兔胃底粘膜cDNA文库,寻找兔胃H,K-ATP酶N端结构域的结合伴侣。我们提取了5个共享超过90%序列同一性的克隆。最长的克隆编码的蛋白质与膜结构域的片段(从Arg-835至Ser-873)以及人和小鼠锚蛋白III的“血影蛋白结合结构域”的主要部分(从Glu-874至Leu-1455)具有高度同一性(分别为96%和96.8%)。我们得出结论,膜和血影蛋白结合域的兔锚蛋白III的兔胃H,K-ATP酶的N-末端结构域的结合伴侣的候选人。为了验证锚定-ATP酶相互作用并测试其特异性,我们在酵母和细菌中产生了这两个结构域,用抗ATP酶抗体共免疫沉淀它们,并通过亲和层析共纯化它们。兔锚蛋白III的序列尚不清楚,这是第一份证明锚蛋白III和H,K-ATP酶相互作用没有中间体的报告。这种相互作用一方面涉及ATP酶的N-末端结构域,另一方面涉及锚蛋白的血影蛋白结合结构域。
We screened a cDNA bank of rabbit gastric fundic mucosa by two-hybrid assays looking for binding partners of the N-terminal domain of the rabbit gastric H,K-ATPase. We extracted five clones sharing more than 90% sequence identity. The longest clone codes for a protein sharing a high identity (96 and 96.8%, respectively) with a fragment of the membrane domain, from Arg-835 to Ser-873, plus the major part of the “spectrin binding domain” going from Glu-874 to Leu-1455 of human and mouse ankyrin III. We conclude that the membrane and spectrin binding domains of the rabbit ankyrin III are candidates for the binding partner of the N-terminal domain of the rabbit gastric H,K-ATPase. To validate the ankyrin-ATPase interaction and to test its specificity, we produced both domains in yeast and bacteria, coimmunoprecipitated them with an anti-ATPase antibody, and copurified them by affinity chromatography. The sequence of rabbit ankyrin III was not known, and this is the first report demonstrating that the ankyrin III and the H,K-ATPase interact with no intermediate. The interaction involves the N-terminal domain of the ATPase on one hand and the spectrin binding domain of the ankyrin on the other.
DOI: 10.1126/science.2672330
发表时间: 1989-08-18
期刊: SCIENCE
影响因子: 56.9
作者:
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DOI: 10.1152/ajpcell.1993.264.1.c63
发表时间: 1993
期刊: The American journal of physiology
影响因子: --
作者:
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期刊: The Journal of biological chemistry
影响因子: --
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发表时间: 1992-11-06
期刊: SCIENCE
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