ISOLATION OF CDNAS FOR PERILIPIN-A AND PERILIPIN-B - SEQUENCE AND EXPRESSION OF LIPID DROPLET-ASSOCIATED PROTEINS OF ADIPOCYTES
ISOLATION OF CDNAS FOR PERILIPIN-A AND PERILIPIN-B - SEQUENCE AND EXPRESSION OF LIPID DROPLET-ASSOCIATED PROTEINS OF ADIPOCYTES
复制标题
DOI：
10.1073/pnas.90.24.12035
复制
发表时间：
1993-12-15
影响因子：
11.1
通讯作者：
KIMMEL, AR
中科院分区：
文献类型：
--
作者：
GREENBERG, AS;EGAN, JJ;KIMMEL, AR
脂肪细胞中主要的cAMP依赖蛋白激酶(A-Kinase)底物是Perilipin，这是一种仅存在于脂质储存液滴表面的蛋白质。利用抗Perilipin血清，我们从大鼠脂肪细胞cDNA表达文库中分离到两类相关的全长编码基因，命名为Perilipin A和B。这两个cDNA来自两个通过差异剪接产生的mRNA物种。预测的mRNAs编码Perilipins A和B，分别为517个氨基酸(56,870 Da)和422个AA(46,420 Da)的蛋白质，它们共享一个共同的406-AA N-末端序列。预测的Perilipin A包含从大鼠脂肪细胞中纯化的62 kDa Perilipin的蛋白水解酶中存在的多肽，以及必要的共识A-激酶磷酸化位点。像Perilipin A一样，B形式在脂肪细胞中表达，并与脂质储存液滴有关。对预测的二级结构的模拟未能揭示紫苏素与脂滴之间坚韧联系的潜在基础。这些蛋白质与另一种已知蛋白质--脂肪细胞分化相关蛋白(Adrp)显示出显著的序列关系(通过105aa几乎等于65%的相似性)。像Perilipins一样，adrp似乎是脂肪细胞特异性的，这表明它们在相关的细胞内途径中相互作用。这里描述的紫苏素A和B的分子探针将允许详细分析它们在脂类代谢中的功能作用(S)。
The major cAMP-dependent protein kinase (A-kinase) substrate in adipocytes is perilipin, a protein found exclusively at the surface of the lipid storage droplets. Using anti-perilipin serum, we have isolated two related classes of full-length coding cDNAs, designated perilipin A and B, from a rat adipocyte cDNA expression library. The two cDNAs derive from two mRNA species that arise by differential splicing. The mRNAs are predicted to encode perilipins A and B, proteins of 517 aa (56,870 Da) and 422 aa (46,420 Da), respectively, which share a common 406-aa N-terminal sequence. The predicted perilipin A contains peptides present in proteolytic digests of the purified 62-kDa form of perilipin from rat adipocytes, as well as the requisite consensus A-kinase phosphorylation sites. Like perilipin A, the B form is expressed in adipocytes and is associated with lipid storage droplets. Modeling of predicted secondary structures fails to reveal an underlying basis for the tenacious association of perilipins with lipid droplets. These proteins exhibit a significant sequence relationship (almost-equal-to 65% similarity through 105 aa) with only one other known protein, the adipocyte differentiation-related protein (ADRP). Like the perilipins, ADRP appears to be adipocyte-specific, which suggests that they interact in a related intracellular pathway. The molecular probes for perilipins A and B described here will permit detailed analyses of their functional role(s) in lipid metabolism.