Hiramatsu T, Matsuo S, Okura T, Yoshida F, Watanabe Y, Sakamoto N
Nihon Jinzo Gakkai Shi. 1989 Feb;31(2):125-34.
Antibodies to a 250,000 molecular weight heparan sulfate proteoglycan (HSPG) core protein purified from PYS-2 cells were prepared in the rabbit. After absorption with laminin, the anti-HSPG serum (AHSPGS) was specific for HSPG by enzyme-linked immunoabsorbent assay (ELISA). Immunoelectron microscopy showed that AHSPGS reacted with the mouse glomerular basement membrane (GBM) or, more precisely, with three HSPG rich layers, i.e., (i) the lamina rara interna in a discrete or interrupted-linear pattern, (ii) the interface between the GBM and the soles of the foot processes, and (iii) the lamina rara externa in a finely discrete pattern. The localization of HSPG in the GBM was very similar to that of the fixed negative charges detected with polyethyleneimine (PEI). When mice were treated with heparitinase, binding of PEI was seen only on the cell surface including the foot process soles (ii) but not in the GBM (i and iii), while the localization of HSPG core protein was unaltered. These results suggest that the localization of the HSPG core protein was correlated with that of the fixed negative charges in the GBM (i and iii). The foot process base (ii) seems to be a complicated zone since the PEI binding in this area was less affected by heparitinase treatment, while this area has been reported to have a lower sialic acid content than the foot process surface facing the urinary space.
从PYS - 2细胞中纯化出分子量为250,000的硫酸乙酰肝素蛋白聚糖（HSPG）核心蛋白，并以此制备兔抗抗体。用层粘连蛋白吸收后，通过酶联免疫吸附测定（ELISA），抗HSPG血清（AHSPGS）对HSPG具有特异性。免疫电子显微镜显示，AHSPGS与小鼠肾小球基底膜（GBM）发生反应，更确切地说，是与富含HSPG的三层结构发生反应，即：（i）内疏松层呈离散或间断线性模式；（ii）GBM与足突底部之间的界面；（iii）外疏松层呈精细离散模式。GBM中HSPG的定位与用聚乙烯亚胺（PEI）检测到的固定负电荷的定位非常相似。当用肝素酶处理小鼠时，PEI的结合仅见于包括足突底部（ii）在内的细胞表面，而不见于GBM（i和iii），而HSPG核心蛋白的定位未改变。这些结果表明，HSPG核心蛋白的定位与GBM（i和iii）中固定负电荷的定位相关。足突基部（ii）似乎是一个复杂的区域，因为该区域的PEI结合受肝素酶处理的影响较小，而据报道该区域的唾液酸含量低于面向尿腔的足突表面。
