Liu Xiaoxi, Liu Jingjing, Lester Joshua D, Pijut Sonja S, Graf Gregory A
Department of Pharmaceutical Sciences, Saha Cardiovascular Research Center, Barnstable Brown Diabetes and Obesity Center, University of Kentucky, Lexington, KY, United States.
Biochem Biophys Res Commun. 2015 Jan 2;456(1):129-34. doi: 10.1016/j.bbrc.2014.11.046. Epub 2014 Nov 24.
ATP-binding cassette transporter D2 (D2) is an ABC half transporter that is thought to promote the transport of very long-chain fatty acyl-CoAs into peroxisomes. Both D2 and peroxisomes increase during adipogenesis. Although peroxisomes are essential to both catabolic and anabolic lipid metabolism, their function, and that of D2, in adipose tissues remain largely unknown. Here, we investigated the D2 localization and the proteome of D2-containing organelles, in adipose tissue. Centrifugation of mouse adipose homogenates generated a fraction enriched with D2, but deficient in peroxisome markers including catalase, PEX19, and ABCD3 (D3). Electron microscopic imaging of this fraction confirmed the presence of D2 protein on an organelle with a dense matrix and a diameter of ∼ 200 nm, the typical structure and size of a microperoxisome. D2 and PEX19 antibodies recognized distinct structures in mouse adipose. Immunoisolation of the D2-containing compartment confirmed the scarcity of PEX19 and proteomic profiling revealed the presence of proteins associated with peroxisome, endoplasmic reticulum (ER), and mitochondria. D2 is localized to a distinct class of peroxisomes that lack many peroxisome proteins, and may associate physically with mitochondria and the ER.
ATP结合盒转运蛋白D2(D2)是一种ABC半转运蛋白,被认为可促进超长链脂肪酰辅酶A进入过氧化物酶体。在脂肪生成过程中,D2和过氧化物酶体均会增加。尽管过氧化物酶体对分解代谢和合成代谢的脂质代谢都至关重要,但其在脂肪组织中的功能以及D2的功能在很大程度上仍不清楚。在此,我们研究了脂肪组织中D2的定位以及含D2细胞器的蛋白质组。对小鼠脂肪匀浆进行离心,得到了一个富含D2但缺乏包括过氧化氢酶、PEX19和ABCD3(D3)在内的过氧化物酶体标记物的组分。对该组分进行电子显微镜成像,证实了在一个具有致密基质且直径约为200nm的细胞器上存在D2蛋白,这是微过氧化物酶体的典型结构和大小。D2和PEX-19抗体识别出小鼠脂肪中的不同结构。对含D2区室的免疫分离证实了PEX19的稀缺性,蛋白质组分析揭示了与过氧化物酶体、内质网(ER)和线粒体相关的蛋白质的存在。D2定位于一类独特的过氧化物酶体,这类过氧化物酶体缺乏许多过氧化物酶体蛋白,并且可能与线粒体和内质网发生物理关联。