Bhat Eijaz Ahmed, Sajjad Nasreena, Sabir Jamal S M, Kamli Majid Rasool, Hakeem Khalid Rehman, Rather Irfan A, Bahieldin Ahmed
Life Science Institute, Zhejiang University, Hangzhou, Zhejiang 310058, PR China.
Department of Biochemistry, University of Kashmir, Hazratbal, Jammu and Kashmir, India.
Saudi J Biol Sci. 2020 Jun;27(6):1562-1565. doi: 10.1016/j.sjbs.2020.03.011. Epub 2020 Mar 13.
The TRAIP interacting protein is known as a negative regulator of TNF-induced-nuclear factor, kappa-light-chain-enhancer of activated B cell (NF-κB) by direct interaction with the adaptor protein TRAF2, which inhibits the function of TRAF2 via the RINGCC domain protein. The TRAIP protein is composed of 469 amino acids with an N-terminal RING motif that is followed by a coiled coil (CC) and leucine zipper domain. TRAIP proteins are critical in programmed cell death, cell proliferation and differentiation, and embryonic development. The critical functions of TRAIP together with the molecular inhibitory mechanism effect of TRAIP have been reported by two different studies and have opened up new research into the field of TRAF biology. In this study, we designed different constructs of the Leucine zipper domain to find the over -expressed construct for further studies. We successfully cloned the C-terminal TRAIP containing the leucine zipper domain. In addition, we have over-expressed and purified the TRAIP LZ for their biochemical characterization.
TRIP相互作用蛋白被认为是肿瘤坏死因子诱导的核因子（即活化B细胞的κ轻链增强子，NF-κB）的负调节因子，它通过与衔接蛋白TRAF2直接相互作用来实现这一功能，而TRAF2通过RINGCC结构域蛋白抑制TRAF2的功能。TRIP蛋白由469个氨基酸组成，其N端有一个RING基序，后面跟着一个卷曲螺旋（CC）和亮氨酸拉链结构域。TRIP蛋白在程序性细胞死亡、细胞增殖和分化以及胚胎发育中起着关键作用。两项不同的研究报道了TRIP的关键功能以及TRIP的分子抑制机制效应，这为TRAF生物学领域开辟了新的研究方向。在本研究中，我们设计了不同的亮氨酸拉链结构域构建体，以找到过表达的构建体用于进一步研究。我们成功克隆了包含亮氨酸拉链结构域的TRIP C端。此外，我们已经对TRIP LZ进行了过表达和纯化，以进行其生化特性分析。
