Sheela Binu, George Genu, Mandagini Geetha, Appukuttan Padinjaradath S
Department of Biochemistry, Sree Chitra Tirunal Institute for Medical Sciences and Technology, Thiruvananthapuram, 695011, India.
Glycoconj J. 2016 Dec;33(6):953-961. doi: 10.1007/s10719-016-9713-z. Epub 2016 Aug 19.
Lipoprotein (a) [Lp(a)] is the dominant lipid in atherosclerotic plaques though it is much less numerous than LDL or HDL in circulation. Molecular mechanism of selective uptake of Lp(a) into macrophages is unclear. Lp(a) was reported to form circulating immune complexes with the IgG-dominated plasma anti-α-galactoside antibody (anti-Gal) using the serine- and threonine-rich peptide sequences ( STPS) on its apo(a) subunit as surrogate ligand but left the other binding site of antibody free. We examined if these monovalent immune complexes could bind to smaller STPS-containing molecules on macrophage surface. Using placental membrane O-glycosylated proteins (PMOP) isolated by lectin affinity chromatography as model it was shown that human cell surface glycoproteins were small enough to occupy both binding sites of anti-Gal since they increased the fluorescence of FITC label at Fc part of anti-Gal and inhibited binding of anti-Gal and Griffonia simplicifolia lectin of similar specificity to immobilized ligands. Pre-incubation with anti-Gal facilitated Lp(a) attachment to macrophages unless anti-Gal-specific sugar was present. Anti-Gal-mediated attachment of apo(a) to macrophages increased with the number of apo(a) subunits. Further, anti-Gal-mediated binding of the same sample of apo(a) increased with the specific activity of anti-Gal sample. Finally binding of anti-Gal and anti-Gal-apo(a) complex to PMOP and macrophages respectively was mostly inhibited by LDL suggesting STPS as major anti-Gal epitopes on the cell surface. Results indicated that circulating Lp(a)-anti-Gal immune complexes anchor on macrophages using STPS-bearing cell surface glycoproteins as ligands and offer a pathway for Lp(a) sequestration into macrophages.
脂蛋白(a)[Lp(a)]是动脉粥样硬化斑块中的主要脂质,尽管其在循环中的数量比低密度脂蛋白(LDL)或高密度脂蛋白(HDL)少得多。Lp(a)被选择性摄取到巨噬细胞中的分子机制尚不清楚。据报道,Lp(a)利用其载脂蛋白(a)[apo(a)]亚基上富含丝氨酸和苏氨酸的肽序列(STPS)作为替代配体,与以IgG为主的血浆抗α-半乳糖苷抗体(抗Gal)形成循环免疫复合物,但抗体的另一个结合位点保持游离。我们研究了这些单价免疫复合物是否能与巨噬细胞表面较小的含STPS分子结合。以通过凝集素亲和层析分离的胎盘膜O-糖基化蛋白(PMOP)为模型,结果表明人类细胞表面糖蛋白足够小,能够占据抗Gal的两个结合位点,因为它们增加了抗Gal Fc部分的FITC标记的荧光,并抑制了抗Gal和具有相似特异性的 Griffonia simplicifolia凝集素与固定化配体的结合。除非存在抗Gal特异性糖,否则与抗Gal预孵育会促进Lp(a)附着于巨噬细胞。抗Gal介导的apo(a)附着于巨噬细胞的作用随apo(a)亚基数量的增加而增强。此外,抗Gal介导的相同apo(a)样品的结合随抗Gal样品的比活性增加而增加。最后,抗Gal和抗Gal-apo(a)复合物分别与PMOP和巨噬细胞的结合大多被LDL抑制,这表明STPS是细胞表面主要的抗Gal表位。结果表明,循环中的Lp(a)-抗Gal免疫复合物以含STPS的细胞表面糖蛋白作为配体锚定在巨噬细胞上,为Lp(a)被巨噬细胞摄取提供了一条途径。