Wang Jianfeng, Wu Gusheng, Miyagi Taeko, Lu Zi-Hua, Ledeen Robert W
Department of Neurology & Neurosciences, New Jersey Medical School, UMDNJ, Newark, New Jersey 07103, USA.
J Neurochem. 2009 Oct;111(2):547-54. doi: 10.1111/j.1471-4159.2009.06339.x. Epub 2009 Aug 17.
Previous reports indicated the presence of both gangliosides and sialidase in the nuclear envelope (NE) of primary neurons and the NG108-15 neural cell line. GM1, one of the major gangliosides of this membrane, was shown to be tightly associated with a sodium-calcium exchanger in the inner membrane of the NE and to potentiate exchanger activity. GD1a was the other major ganglioside detected in the NE and, like GM1, occurs in both inner and outer membranes. A subsequent report indicated the presence of sialidase activity in the NE without specification as to which of the two membranes express it. The present study was undertaken to determine the nature and locus of this activity within the NE of two cell lines: NG108-15 and SH-SY5Y. Western blot analysis of the separated membranes revealed occurrence of Neu3 in the inner membrane and Neu1 in the outer membrane of the NE. Moreover, sialidase activity at both sites was shown capable of catalyzing conversion of endogenous GD1a to GM1.
先前的报告表明,原代神经元和NG108 - 15神经细胞系的核膜(NE)中存在神经节苷脂和唾液酸酶。GM1是该膜的主要神经节苷脂之一,已证明它与核膜内膜中的钠钙交换器紧密相关,并能增强交换器活性。GD1a是在核膜中检测到的另一种主要神经节苷脂,与GM1一样,存在于内膜和外膜中。随后的一份报告表明核膜中存在唾液酸酶活性,但未指明是哪一层膜表达该酶。本研究旨在确定两种细胞系(NG108 - 15和SH - SY5Y)核膜内这种活性的性质和位置。对分离出的膜进行蛋白质免疫印迹分析显示,Neu3存在于核膜的内膜中,Neu1存在于核膜的外膜中。此外,两个部位的唾液酸酶活性均显示能够催化内源性GD1a转化为GM1。