Hussain Zahir, Uyama Toru, Kawai Katsuhisa, Rahman Iffat Ara Sonia, Tsuboi Kazuhito, Araki Nobukazu, Ueda Natsuo
Department of Biochemistry Kagawa University School of Medicine, Miki, Kagawa 761-0793, Japan.
Department of Histology and Cell Biology, Kagawa University School of Medicine, Miki, Kagawa 761-0793, Japan.
J Lipid Res. 2016 Nov;57(11):2051-2060. doi: 10.1194/jlr.M071290. Epub 2016 Sep 13.
N-Acylphosphatidylethanolamines (NAPEs) are a class of glycerophospholipids, which are known as precursors for different bioactive N-acylethanolamines. We previously reported that phospholipase A/acyltransferase-1 (PLAAT-1), which was originally found in mammals as a tumor suppressor, catalyzes N-acylation of phosphatidylethanolamines to form NAPEs. However, recent online database suggested the presence of an uncharacterized isoform of PLAAT-1 with an extra sequence at the N terminus. In the present study, we examined the occurrence, intracellular localization, and catalytic properties of this longer isoform, as well as the original shorter isoform from humans and mice. Our results showed that human tissues express the longer isoform but not the short isoform at all. In contrast, mice expressed both isoforms with different tissue distribution. Unlike the cytoplasmic localization of the shorter isoform, the long isoform was found in both cytoplasm and nucleus, inferring that the extra sequence harbors a nuclear localization signal. As assayed with purified proteins, neither isoform required calcium for full activity. Moreover, the overexpression of each isoform remarkably increased cellular NAPE levels. These results conclude that the new long isoform of PLAAT-1 is a calcium-independent N-acyltransferase existing in both cytoplasm and nucleus and suggest a possible formation of NAPEs in various membrane structures including nuclear membrane. J. Lipid Res 2016. 57: 2051-2060.
N-酰基磷脂酰乙醇胺(NAPEs)是一类甘油磷脂,被认为是不同生物活性N-酰基乙醇胺的前体。我们之前报道过,磷脂酶A/酰基转移酶-1(PLAAT-1)最初作为一种肿瘤抑制因子在哺乳动物中被发现,它催化磷脂酰乙醇胺的N-酰化反应以形成NAPEs。然而,最近的在线数据库显示存在一种未被鉴定的PLAAT-1同工型,其N端有一段额外序列。在本研究中,我们检测了这种较长同工型以及人和小鼠中原始较短同工型的存在情况、细胞内定位和催化特性。我们的结果表明,人类组织表达较长同工型,但完全不表达短同工型。相反,小鼠表达这两种同工型,且组织分布不同。与较短同工型的胞质定位不同,较长同工型在胞质和细胞核中均有发现,这表明额外序列含有一个核定位信号。用纯化蛋白进行检测时,两种同工型的充分活性均不需要钙。此外,每种同工型的过表达均显著提高了细胞内NAPEs水平。这些结果表明,新发现的PLAAT-1长同工型是一种存在于胞质和细胞核中的不依赖钙的N-酰基转移酶,并提示在包括核膜在内的各种膜结构中可能形成NAPEs。《脂质研究杂志》2016年。57: 2051 - 2060。