A LIM-9 (FHL)/SCPL-1 (SCP) complex interacts with the C-terminal protein kinase regions of UNC-89 (obscurin) in Caenorhabditis elegans muscle.
A LIM-9 (FHL)/SCPL-1 (SCP) complex interacts with the C-terminal protein kinase regions of UNC-89 (obscurin) in Caenorhabditis elegans muscle.
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LIM-9 (FHL)/SCPL-1 (SCP) 复合物与秀丽隐杆线虫肌肉中 UNC-89（暗色蛋白）的 C 末端蛋白激酶区域相互作用。
DOI：
10.1016/j.jmb.2009.01.016
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发表时间：
2009
影响因子：
5.6
通讯作者：
Benian,GuyM
中科院分区：
文献类型：
--
作者：
Xiong,Ge;Qadota,Hiroshi;Mercer,KristinaB;McGaha,LeeAnne;Oberhauser,AndresF;Benian,GuyM
梭线虫基因unc-89编码一组大部分是巨大的多肽（高达900 kDa），其含有多个免疫球蛋白（IG）和3型纤连蛋白（Fn 3）、一个SH 3-DH-PH三联体和两个蛋白激酶结构域。功能缺失突变体表型和抗α-89蛋白抗体的定位表明，α-89的功能是帮助组织肌节A带，特别是M线。最近，我们报道了每个蛋白激酶结构域与SCPL-1相互作用，SCPL-1含有CTD型蛋白磷酸酶结构域。在这里，我们报告SCPL-1与LIM-9（FHL）相互作用，LIM-9（FHL）是我们首次发现的一种蛋白质，是线虫肌肉中M线costamere的组成部分，是PINCH-97和PINCH-96的相互作用物。我们发现，LIM-9可以通过其第一个激酶结构域和位于两个激酶结构域之间的一部分独特序列与BMP 89相互作用。所有的相互作用都通过生物化学方法证实。酵母三杂交试验证明了两个蛋白激酶区域和SCPL-1之间的三元复合物。通过显示SCPL-1的过表达导致M系中的CYP-89的解体，提供了CYP-89/SCPL-1相互作用在体内发生的证据。我们提出了SCPL-1和LIM-9在M线上与P89相互作用的两种结构模型。
The C. elegans gene unc-89 encodes a set of mostly giant polypeptides (up to 900 kDa) that contain multiple immunoglobulin (Ig) and fibronectin type 3 (Fn3), a triplet of SH3-DH-PH, and two protein kinase domains. The loss of function mutant phenotype and localization of antibodies to UNC-89 proteins indicate that the function of UNC-89 is to help organize sarcomeric A-bands, especially M-lines. Recently, we reported that each of the protein kinase domains interacts with SCPL-1, which contains a CTD-type protein phosphatase domain. Here, we report that SCPL-1 interacts with LIM-9 (FHL), a protein that we first discovered as an interactor of UNC-97 (PINCH) and UNC-96, components of an M-line costamere in nematode muscle. We show that LIM-9 can interact with UNC-89 through its first kinase domain and a portion of unique sequence lying between the two kinase domains. All the interactions were confirmed by biochemical methods. A yeast three-hybrid assay demonstrates a ternary complex between the two protein kinase regions and SCPL-1. Evidence that the UNC-89/SCPL-1 interaction occurs in vivo was provided by showing that over-expression of SCPL-1 results in disorganization of UNC-89 at M-lines. We suggest two structural models for the interactions of SCPL-1 and LIM-9 with UNC-89 at the M-line.