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由小球藻病毒编码的小钾离子通道蛋白。

Small potassium ion channel proteins encoded by chlorella viruses.

作者信息

Kang Ming, Moroni Anna, Gazzarrini Sabrina, DiFrancesco Dario, Thiel Gerhard, Severino Maria, Van Etten James L

机构信息

Department of Plant Pathology, University of Nebraska, Lincoln, NE 68583-0722, USA.

出版信息

Proc Natl Acad Sci U S A. 2004 Apr 13;101(15):5318-24. doi: 10.1073/pnas.0307824100. Epub 2004 Feb 4.

Abstract

Kcv, a 94-aa protein encoded by Paramecium bursaria chlorella virus 1, is the smallest known protein to form a functional potassium ion channel and basically corresponds to the "pore module" of potassium channels. Both viral replication and channel activity are inhibited by the ion channel blockers barium and amantadine but not by cesium. Genes encoding Kcv-like proteins were isolated from 40 additional chlorella viruses. Differences in 16 of the 94 amino acids were detected, producing six Kcv-like proteins with amino acid substitutions occurring in most of the functional domains of the protein (N terminus, transmembrane 1, pore helix, selectivity filter, and transmembrane 2). The six proteins form functional potassium selective channels in Xenopus oocytes with different properties including altered current kinetics and inhibition by cesium. The amino acid changes together with the different properties observed in the six Kcv-like channels will be used to guide site-directed mutations, either singularly or in combination, to identify key amino acids that confer specific properties to Kcv.

摘要

Kcv是由草履虫小球藻病毒1编码的一种94个氨基酸的蛋白质,是已知能形成功能性钾离子通道的最小蛋白质,基本对应于钾通道的“孔模块”。病毒复制和通道活性均受到离子通道阻滞剂钡和金刚烷的抑制,但不受铯的抑制。从另外40种小球藻病毒中分离出了编码Kcv样蛋白的基因。在94个氨基酸中的16个氨基酸上检测到差异,产生了6种Kcv样蛋白,其氨基酸取代发生在该蛋白的大部分功能域(N端、跨膜1、孔螺旋、选择性过滤器和跨膜2)。这6种蛋白在非洲爪蟾卵母细胞中形成具有不同特性的功能性钾选择性通道,包括改变的电流动力学和被铯抑制。这6种Kcv样通道中观察到的氨基酸变化以及不同特性将用于指导单点或组合的定点突变,以鉴定赋予Kcv特定特性的关键氨基酸。

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