重组a群链球菌磷酸甘油醛脱氢酶与脂蛋白(a)的相互作用.pdfVIP

摘 要 三磷酸甘油醛脱氢酶(GAPDH)是A群链球菌(group A Streptococcus, GAS)表面的 纤溶酶原(Plg)受体之一，其C末端的赖氨酸残基可以与Plg 的赖氨酸结合位点(LBS)相 结合。脂蛋白(a)[Lp(a)] 中的载脂蛋白(a)[Apo(a)]与Plg有很高的同源性，也含有LBS ， 因此本实验室提出了Lp(a)可能与GAS表面的Plg受体相结合，进而可能竞争性抑制 GAS与Plg结合的假说。而且本实验室已经证明了GAS表面的烯醇化酶能够与Lp(a)发 生特异性结合。 为了更进一步探索Lp(a)是否与其它的GAS表面Plg受体结合，本研究克隆了GAS 的GAPDH及其C末端敲除赖氨酸残基的突变体GAPDHΔ345基因，酶切后将其连接到 表达载体pASK-IBA37 中，并在大肠杆菌BL-21 中表达了这两个重组蛋白(rGAPDH和 rGAPDHΔ345) 。通过酶联免疫吸附试验(ELISA)对rGAPDH和rGAPDHΔ345与Lp(a) 的 相互作用机制进行了研究。结果表明rGAPDHΔ345与Lp(a) 的结合值比rGAPDH显著降 低，而且一定浓度的赖氨酸类似物6-氨基己酸(EACA) 可以抑制这种结合，说明 rGAPDH及rGAPDHΔ345与Lp(a)通过LBS发生特异性结合。另外，Lp(a)对rGAPDH与 Plg 的相互作用也有明显的抑制。 关键词：三磷酸甘油醛脱氢酶；A群链球菌；纤溶酶原；脂蛋白(a) ；赖氨酸结合位点 The Interaction Between Lipoprotein(a) and Recombinant Glyceraldehyde-3-phosphate Dehydrogenase derived from Group A Streptococcus Abstract Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is one of plasminogen (Plg) receptors on the surface of group A streptococcus (GAS). Plg through its lysine binding sites (LBS) binds to GAPDH. Apolipoprotein(a) [Apo(a)], which is one component of Lipoprotein(a) [Lp(a)], has a high homology with Plg. Therefore, we previously postulated that Lp(a) might bind to Plg receptor on the surface of GAS, subsequently competitively inhibiting the interaction between GAS and Plg. Moreover, this hypothesis was demonstrated by Streptococcal surface enolase-Lp(a) interaction assay in our lab. To further test our hypothesis, the GAPDH gene and its C-terminal lysine residues-truncated fragment variant of GAS was cloned into pASK-IBA37 respectively, and the recombinant GAPDH (rGAPDH) and GAPDHΔ345 (rGAPDHΔ345) were expressed in E. coli BL21. The interaction between rGAPDH or rGAPDHΔ345 and Lp(a) was investigated b