Sud'ina G F, Brock T G, Pushkareva M A, Galkina S I, Turutin D V, Peters-Golden M, Ullrich V
A.N. Belozersky Institute of Physico-Chemical Biology, Moscow State University, Moscow 119899, Russia.
Biochem J. 2001 Nov 1;359(Pt 3):621-9. doi: 10.1042/0264-6021:3590621.
Sulphatides are sulphate esters of galactocerebrosides that are present on the surfaces of many cell types and act as specific ligands to selectins. The present study was undertaken to investigate the effect of sulphatides on polymorphonuclear granulocyte (PMN) attachment, spreading and 5-lipoxygenase (5-LO) metabolism. Sulphatides, but not non-sulphated galactocerebrosides, dose-dependently enhanced attachment to collagen, as measured by the myeloperoxidase assay. Studies with blocking antibodies indicated that the increased attachment was mediated by CD11b/CD18 (Mac-1) beta 2 integrin. Scanning electron microscopy indicated that sulphatides also greatly enhanced the degree of cell spreading. In PMNs treated in suspension, sulphatides had no effect on the ionophore A23187-stimulated release of arachidonic acid and the synthesis of 5-LO metabolites. In contrast, in PMNs attached to collagen, the enzymic conversion of arachidonic acid by 5-LO was inhibited by sulphatides. Inhibition of 5-LO metabolism by sulphatides was observed even in the presence of exogenous substrate, suggesting that sulphatides directly inhibited 5-LO action. Consistent with this, sulphatides interfered with ionophore-induced translocation of the 5-LO to the nuclear envelope. Substances competing with sulphatide binding to cells, like dextran sulphate, or a strong inhibitor of cell spreading, like the actin-polymerizing agent jasplakinolide, prevented the effects of sulphatides on PMN attachment and spreading and leukotriene synthesis. We conclude that shape changes occurring in response to sulphatides specifically impair PMN leukotriene synthesis by inhibiting translocation of 5-LO.
硫脂是半乳糖脑苷脂的硫酸酯,存在于多种细胞表面,作为选择素的特异性配体。本研究旨在探讨硫脂对多形核粒细胞(PMN)黏附、铺展及5-脂氧合酶(5-LO)代谢的影响。通过髓过氧化物酶测定法检测,硫脂而非非硫酸化的半乳糖脑苷脂能剂量依赖性地增强对胶原蛋白的黏附。用阻断抗体进行的研究表明,黏附增加是由CD11b/CD18(Mac-1)β2整合素介导的。扫描电子显微镜显示,硫脂也极大地增强了细胞铺展程度。在悬浮处理的PMN中,硫脂对离子载体A23187刺激的花生四烯酸释放及5-LO代谢产物的合成没有影响。相反,在黏附于胶原蛋白的PMN中,硫脂抑制了5-LO对花生四烯酸的酶促转化。即使存在外源性底物,硫脂对5-LO代谢的抑制作用仍可观察到,这表明硫脂直接抑制了5-LO的作用。与此一致的是,硫脂干扰了离子载体诱导的5-LO向核膜的转位。与硫脂竞争结合细胞的物质,如硫酸葡聚糖,或细胞铺展的强抑制剂,如肌动蛋白聚合剂茉莉酮酸酯,可阻止硫脂对PMN黏附、铺展及白三烯合成的影响。我们得出结论,硫脂引起的形状变化通过抑制5-LO的转位特异性损害PMN白三烯的合成。