Is outer arm dynein intermediate chain 1 multifunctional?
Is outer arm dynein intermediate chain 1 multifunctional?
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DOI：
10.1091/mbc.7.12.1895
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发表时间：
1996-12-01
影响因子：
3.3
通讯作者：
Mohri, H
中科院分区：
文献类型：
--
作者：
Ogawa, K;Takai, H;Mohri, H
海胆精子轴丝的外臂动力蛋白含有三条中间链（IC 1、IC 2和IC 3; M（r）分别为128，000、98，000和74，000）。IC 2和IC 3是WD家族的成员; WD基序负责蛋白质-蛋白质相互作用。我们在这里描述的IC 1的分子克隆。IC 1具有独特的一级结构，N端部分与硫氧还蛋白序列同源，中间部分由三个与核苷二磷酸激酶序列同源的重复序列组成，C端部分含有高比例的带负电荷的谷氨酸残基。因此，IC 1是一种不同于IC 2和IC 3的新型动力蛋白中间链，可能是一种多功能蛋白。IC 1的硫氧还蛋白相关部分与两个氧化还原活性衣原体轻链比硫氧还蛋白更密切相关。针对在细菌中表达为His标记蛋白的IC 1的N-末端和中间结构域制备抗体。这些抗体与来自远亲物种的一些动力蛋白多肽（IC 1的潜在同系物）发生交叉反应。我们认为这三条中间链是精子外臂动力蛋白的基本核心单元，因为它们普遍存在。从海胆和远亲物种的重组硫氧还蛋白相关的部分IC 1和外臂动力蛋白特异性结合，并从苯胂氧化物亲和柱与2-巯基乙醇洗脱，表明它们含有邻位二硫醇能够进行可逆的氧化/还原。
The outer arm dynein of sea urchin sperm axoneme contains three intermediate chains (IC1, IC2, and IC3; M(r) 128,000, 98,000, and 74,000, respectively). IC2 and IC3 are members of the WD family; the WD motif is responsible for a protein-protein interaction. We describe here the molecular cloning of IC1. IC1 has a unique primary structure, the N-terminal part is homologous to the sequence of thioredoxin, the middle part consists of three repetitive sequences homologous to the sequence of nucleoside diphosphate kinase, and the C-terminal part contains a high proportion of negatively charged glutamic acid residues. Thus, IC1 is a novel dynein intermediate chain distinct from IC2 and IC3 and may be a multifunctional protein. The thioredoxin-related part of IC1 is more closely related to those of two redox-active Chlamydomonas light chains than thioredoxin. Antibodies were prepared against the N-terminal and middle domains of IC1 expressed as His-tagged proteins in bacteria. These antibodies cross-reacted with some dynein polypeptides (potential homologues of IC1) from distantly related species. We propose here that the three intermediate chains are the basic core units of sperm outer arm dynein because of their ubiquitous existence. The recombinant thioredoxin-related part of IC1 and outer arm dyneins from sea urchin and distantly related species were specifically bound to and eluted from a phenylarsine oxide affinity column with 2-mercaptoethanol, indicating that they contain vicinal dithiols competent to undergo reversible oxidation/reduction.