Proteomic investigation of the interactome of FMNL1 in hematopoietic cells unveils a role in calcium-dependent membrane plasticity
Proteomic investigation of the interactome of FMNL1 in hematopoietic cells unveils a role in calcium-dependent membrane plasticity
复制标题
DOI：
10.1016/j.jprot.2012.11.015
复制
发表时间：
2013-01-14
影响因子：
3.3
通讯作者：
Krackhardt, Angela M.
中科院分区：
文献类型：
--
作者：
Han, Yanan;Yu, Guangchuang;Krackhardt, Angela M.
FMNL 1是一种在造血细胞中高度表达、在白血病和多种转化细胞系中过表达的formin相关蛋白。已描述其在造血细胞的多种功能中发挥作用，例如巨噬细胞的吞噬作用以及T细胞的极化和细胞毒性。然而，FMNL 1在这些过程中的具体作用还没有得到澄清，在初级造血细胞中的相互作用伴侣的调节从未被研究过。我们进行了一个蛋白质组学筛选的FMNL 1在原代造血细胞中的相互作用组的调查，从而确定了一些相互作用的合作伙伴。考虑语义相似性的生物信息学分析表明，AHNAK 1是FMNL 1的重要相互作用伙伴。我们证实AHNAK 1作为FMNL 1在不同造血细胞中的一般结合伴侣，并证明FMNL 1的N-末端部分与AHNAK 1的C-末端结合。此外，我们表明，组成型激活形式的FMNL 1（FMNL 1 γ）诱导AHNAK 1的定位到细胞膜。最后，我们提供的证据表明，过表达或敲低FMNL 1的离子霉素介导的激活后，不同的细胞系和原代细胞的能力钙内流的影响。(C)2012爱思唯尔有限公司版权所有。
Formin-like 1 (FMNL1) is a formin-related protein highly expressed in hematopoietic cells and overexpressed in leukemias as well as diverse transformed cell lines. It has been described to play a role in diverse functions of hematopoietic cells such as phagocytosis of macrophages as well as polarization and cytotoxicity of T cells. However, the specific role of FMNL1 in these processes has not been clarified yet and regulation by interaction partners in primary hematopoietic cells has never been investigated. We performed a proteomic screen for investigation of the interactome of FMNL1 in primary hematopoietic cells resulting in the identification of a number of interaction partners. Bioinformatic analysis considering semantic similarity suggested the giant protein AHNAK1 to be an essential interaction partner of FMNL1. We confirmed AHNAK1 as a general binding partner for FMNL1 in diverse hematopoietic cells and demonstrate that the N-terminal part of FMNL1 binds to the C-terminus of AHNAK1. Moreover, we show that the constitutively activated form of FMNL1 (FMNL1 gamma) induces localization of AHNAK1 to the cell membrane. Finally, we provide evidence that overexpression or knock down of FMNL1 has an impact on the capacitative calcium influx after ionomycin-mediated activation of diverse cell lines and primary cells. (C) 2012 Elsevier B.V. All rights reserved.