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果蝇驱动蛋白轻链。一级结构及其与驱动蛋白重链的相互作用。

The Drosophila kinesin light chain. Primary structure and interaction with kinesin heavy chain.

作者信息

Gauger A K, Goldstein L S

机构信息

Department of Cell and Developmental Biology, Harvard University, Cambridge, Massachusetts 02138.

出版信息

J Biol Chem. 1993 Jun 25;268(18):13657-66.

PMID:8514798
Abstract

Kinesin light chain (KLC) complexes with the kinesin heavy chain (KHC) to form native kinesin. Proposed functions of KLC include coupling of cargo to KHC or modulation of KHC ATPase activity. In this paper we use the KHC tail, which binds specifically to KLC in blot overlays, as a probe to clone a cDNA encoding KLC from a Drosophila expression library. The identified clone encodes a protein with 70% amino acid identity to rat KLC. Drosophila KLC is predicted to form an alpha-helical coiled-coil between residues 34 and 129, followed by five imperfect tandem repeats of unknown function and a sixth shorter motif. These repeats are highly conserved across species. The Drosophila KLC gene is located at 69D on the third chromosome and is widely expressed, with 1.8-kb transcripts in most tissues, and slightly smaller transcripts in gonads. Finally, we present evidence that the heptad repeats of KLC are required for interaction with the KHC tail. Since the KHC tail used in our assay includes about 20 heptad repeats, this result suggests that KHC and KLC interact via coiled-coils. Such an interaction could provide stability to the KHC-KLC complex in vivo.

摘要

驱动蛋白轻链(KLC)与驱动蛋白重链(KHC)结合形成天然驱动蛋白。KLC的推测功能包括将货物与KHC偶联或调节KHC的ATP酶活性。在本文中,我们使用在印迹覆盖物中与KLC特异性结合的KHC尾部作为探针,从果蝇表达文库中克隆编码KLC的cDNA。鉴定出的克隆编码一种与大鼠KLC具有70%氨基酸同一性的蛋白质。果蝇KLC预计在第34至129位残基之间形成α螺旋卷曲螺旋结构,随后是五个功能未知的不完全串联重复序列和第六个较短的基序。这些重复序列在物种间高度保守。果蝇KLC基因位于第三条染色体的69D处,广泛表达,大多数组织中有1.8kb的转录本,性腺中的转录本略小。最后,我们提供证据表明KLC的七肽重复序列是与KHC尾部相互作用所必需的。由于我们实验中使用的KHC尾部包含约20个七肽重复序列,这一结果表明KHC和KLC通过卷曲螺旋相互作用。这种相互作用可以在体内为KHC - KLC复合物提供稳定性。

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