Popova N V, Plotnikov A N, Ziganshin R Kh, Deyev I E, Petrenko A G
Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow 117991, Russia.
Biochemistry (Mosc). 2008 Jun;73(6):644-51. doi: 10.1134/s0006297908060035.
Calcium-independent receptor of latrotoxin (CIRL) is an orphan heptahelical receptor implicated in regulation of exocytosis. To characterize molecular mechanisms of CIRL functioning, we searched for its intracellular partners using the yeast two-hybrid SR system with the cytoplasmic C-terminal fragment of CIRL as bait. One of the interacting proteins was identified as TRIP8b, a putative cytosolic adapter protein with multiple tetratricopeptide repeats. To understand functional significance of CIRL-TRIP8b interaction, we further isolated TRIP8b-interacting proteins by affinity chromatography of brain extracts on immobilized recombinant TRIP8b. Sixteen proteins were identified by mass spectrometry in the purified preparations. Clathrin and subunits of AP2 complex appeared to be the major TRIP8b-interacting proteins. Our data suggest a role of TRIP8b in receptor-mediated endocytosis.
无钙黑寡妇毒素受体（CIRL）是一种参与调节胞吐作用的孤儿七螺旋受体。为了阐明CIRL发挥功能的分子机制，我们以CIRL的胞质C末端片段为诱饵，利用酵母双杂交SR系统寻找其细胞内相互作用蛋白。其中一种相互作用蛋白被鉴定为TRIP8b，它是一种具有多个四肽重复序列的假定胞质衔接蛋白。为了了解CIRL-TRIP8b相互作用的功能意义，我们通过固定化重组TRIP8b对脑提取物进行亲和层析，进一步分离出与TRIP8b相互作用的蛋白。通过质谱分析在纯化产物中鉴定出16种蛋白。网格蛋白和AP2复合物亚基似乎是与TRIP8b相互作用的主要蛋白。我们的数据表明TRIP8b在受体介导的内吞作用中发挥作用。
