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首页> 外文期刊>Archives of Biochemistry and Biophysics >Expression, regulation and functional assessment of the 80 amino acid Small Adipocyte Factor 1 (Smaf1) protein in adipocytes
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Expression, regulation and functional assessment of the 80 amino acid Small Adipocyte Factor 1 (Smaf1) protein in adipocytes

机译:脂肪细胞中80个氨基酸的小脂肪细胞因子1(Smaf1)蛋白的表达,调控和功能评估

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The gene for Small Adipocyte Factor 1, Smaf1 (also known as adipogenin, ADIG), encodes a similar to 600 base transcript that is highly upregulated during 3T3-L1 in vitro adipogenesis and markedly enriched in adipose tissues. Based on the lack of an obvious open reading frame in the Smaf1 transcript, it is not known if the Smaf1 gene is protein coding or non-coding RNA. Using a peptide from a putative open reading frame of Smaf1 as antigen, we generated antibodies for western analysis. Our studies prove that Smaf1 encodes an adipose-enriched protein which in western blot analysis migrates at similar to 10 kDa. Rapid induction of Smaf1 protein occurs during in vitro adipogenesis and its expression in 3T3-L1 adipocytes is positively regulated by insulin and glucose. Moreover, siRNA studies reveal that expression of Smaf1 in adipocytes is wholly dependent on PPAR gamma. On the other hand, use of siRNA for Smaf1 to nearly abolish its protein expression in adipocytes revealed that Smaf1 does not have a major role in adipocyte triglyceride accumulation, lipolysis or insulin-stimulated pAkt induction. However, immunolocalization studies using HA-tagged Smaf1 reveal enrichment at adipocyte lipid droplets. Together our findings show that Smaf1 is a novel small protein endogenous to adipocytes and that Smaf1 expression is closely tied to PPAR gamma-mediated signals and the adipocyte phenotype. (C) 2015 Elsevier Inc. All rights reserved.
机译:小脂肪细胞因子1 Smaf1(也称为脂肪生成素,ADIG)的基因编码类似于600个碱基的转录物,该转录物在3T3-L1体外脂肪生成过程中高度上调,并在脂肪组织中显着富集。由于Smaf1转录物中缺乏明显的开放阅读框,因此尚不清楚Smaf1基因是编码蛋白质还是非编码RNA。使用推定的Smaf1开放阅读框的肽作为抗原,我们生成了用于Western分析的抗体。我们的研究证明,Smaf1编码一种富含脂肪的蛋白质,在蛋白质印迹分析中其迁移速率接近10 kDa。 Smaf1蛋白的快速诱导发生在体外脂肪生成过程中,其在3T3-L1脂肪细胞中的表达受到胰岛素和葡萄糖的正调控。此外,siRNA研究表明,脂肪细胞中Smaf1的表达完全依赖于PPARγ。另一方面,将siRNA用于Smaf1几乎消除了其在脂肪细胞中的蛋白质表达,这表明Smaf1在脂肪细胞甘油三酸酯的积累,脂解作用或胰岛素刺激的pAkt诱导中没有主要作用。但是,使用HA标记的Smaf1进行的免疫定位研究表明,脂肪细胞脂滴处富集。在一起我们的发现表明,Smaf1是脂肪细胞内源性的一种新型小蛋白,Smaf1的表达与PPARγ介导的信号和脂肪细胞的表型密切相关。 (C)2015 Elsevier Inc.保留所有权利。

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