Paensuwan Pussadee, Ngoenkam Jatuporn, Khamsri Boonruang, Preechanukul Kanlaya, Sanguansermsri Donruedee, Pongcharoen Sutatip
Department of Microbiology and Parasitology, Faculty of Medical Science; Naresuan University, Pitsanulok 65000 Thailand.
Asian Pac J Allergy Immunol. 2015 Sep;33(3):189-95. doi: 10.12932/AP0544.33.3.2015.
The engagement of the T cell receptor (TCR)-CD3 complex induces the formation of multiple signalling complexes, which are required for actin cytoskeletal rearrangement. The Wiskott-Aldrich syndrome protein (WASp) is a key regulator of actin polymerization that is recruited to the TCR activation site. Since WASp is a binding partner of adaptor protein Nck, which is recruited directly to the TCR CD3? subunit upon TCR ligation, therefore we proposed that the direct recruitment of Nck to TCR-CD3 may also bring WASp directly to TCR-CD3.
The aim of this present study was to assess the distribution of WASp, in relation to Nck, to the TCR-CD3ε complex.
Jurkat T cells were stimulated with anti-TCR antibody and then the cell lysates were immunoprecipitated with anti-CD3 antibody before immunoblotting with antibodies specific to WASp, Nck1, Nck2, SLP-76 and CD3ε molecules.
WASp was recruited to SLP-76 and also directly to the TCR-CD3 complex upon TCR triggering. The inducible recruitment of WASp to the TCR-CD3 complex is partially dependent of tyrosine phosphorylation.
The present findings provide an alternative mechanism of WASp recruitment to the site of TCR activation that may be involved in recruitment of Nck.
T细胞受体（TCR）-CD3复合物的结合诱导多种信号复合物的形成，这是肌动蛋白细胞骨架重排所必需的。威斯科特-奥尔德里奇综合征蛋白（WASp）是肌动蛋白聚合的关键调节因子，被招募到TCR激活位点。由于WASp是衔接蛋白Nck的结合伙伴，Nck在TCR连接时直接被招募到TCR CD3ε亚基上，因此我们推测Nck直接招募到TCR-CD3也可能将WASp直接带到TCR-CD3。
本研究旨在评估WASp与Nck相关的在TCR-CD3ε复合物中的分布情况。
用抗TCR抗体刺激Jurkat T细胞，然后用抗CD3抗体对细胞裂解物进行免疫沉淀，之后用针对WASp、Nck1、Nck2、SLP-76和CD3ε分子的抗体进行免疫印迹。
在TCR触发时，WASp被招募到SLP-76，也直接被招募到TCR-CD3复合物。WASp可诱导性地招募到TCR-CD3复合物部分依赖于酪氨酸磷酸化。
本研究结果提供了一种WASp被招募到TCR激活位点的替代机制，这可能与Nck的招募有关。
