Shimoda Yoko, Okada Shuichi, Yamada Eijiro, Pessin Jeffrey E, Yamada Masanobu
Department of Medicine and Molecular Science (Y.S., S.O., E.Y., M.Y.), Gunma University Graduate School of Medicine, Gunma 371-8511, Japan; and Departments of Medicine and Molecular Pharmacology (J.E.P.), Albert Einstein College of Medicine, Bronx, New York 10461.
Endocrinology. 2015 Oct;156(10):3548-58. doi: 10.1210/en.2015-1120. Epub 2015 Jul 22.
Tctex1d2 (Tctex1 domain containing 2) is an open reading frame that encodes for a functionally unknown protein that contains a Tctex1 domain found in dynein light chain family members. Examination of gene expression during adipogenesis demonstrated a marked increase in Tctex1d2 protein expression that was essentially undetectable in preadipocytes and markedly induced during 3T3-L1 adipocyte differentiation. Tctex1d2 overexpression significantly inhibited insulin-stimulated glucose transporter 4 (GLUT4) translocation and 2-deoxyglucose uptake. In contrast, Tctex1d2 knockdown significantly increased insulin-stimulated GLUT4 translocation and 2-deoxyglucose uptake. However, acute insulin stimulation (up to 30 min) in 3T3-L1 adipocytes with overexpression or knockdown of Tctex1d2 had no effect on Akt phosphorylation, a critical signal transduction target required for GLUT4 translocation. Although overexpression of Tctex1d2 had no significant effect on GLUT4 internalization, Tctex1d2 was found to associate with syntaxin 4 in an insulin-dependent manner and inhibit Doc2b binding to syntaxin 4. In addition, glucose-dependent insulinotropic polypeptide rescued the Tctex1d2 inhibition of insulin-stimulated GLUT4 translocation by suppressing the Tctex1d2-syntaxin 4 interaction and increasing Doc2b-Synatxin4 interactions. Taking these results together, we hypothesized that Tctex1d2 is a novel syntaxin 4 binding protein that functions as a negative regulator of GLUT4 plasma membrane translocation through inhibition of the Doc2b-syntaxin 4 interaction.
Tctex1d2(含Tctex1结构域2)是一个开放阅读框,编码一种功能未知的蛋白质,该蛋白质含有动力蛋白轻链家族成员中发现的Tctex1结构域。对脂肪生成过程中基因表达的检测表明,Tctex1d2蛋白表达显著增加,在前脂肪细胞中基本检测不到,而在3T3-L1脂肪细胞分化过程中显著诱导表达。Tctex1d2过表达显著抑制胰岛素刺激的葡萄糖转运蛋白4(GLUT4)易位和2-脱氧葡萄糖摄取。相反,Tctex1d2基因敲低显著增加胰岛素刺激的GLUT4易位和2-脱氧葡萄糖摄取。然而,在过表达或敲低Tctex1d2的3T3-L1脂肪细胞中,急性胰岛素刺激(长达30分钟)对Akt磷酸化没有影响,Akt磷酸化是GLUT4易位所需的关键信号转导靶点。虽然Tctex1d2过表达对GLUT4内化没有显著影响,但发现Tctex1d2以胰岛素依赖的方式与Syntaxin 4结合,并抑制Doc2b与Syntaxin 4的结合。此外,葡萄糖依赖性促胰岛素多肽通过抑制Tctex1d2-Syntaxin 4相互作用并增加Doc2b-Syntaxin4相互作用,挽救了Tctex1d2对胰岛素刺激的GLUT4易位的抑制作用。综合这些结果,我们推测Tctex1d2是一种新型的Syntaxin 4结合蛋白,通过抑制Doc2b-Syntaxin 4相互作用,作为GLUT4质膜易位的负调节因子发挥作用。