Millis A J, Hoyle M, Kent L
J Cell Physiol. 1986 Jun;127(3):366-72. doi: 10.1002/jcp.1041270304.
In vitro, high density monolayer cultures of vascular smooth muscle cells can be induced to form multicellular nodules. The nodular cells appear to be morphologically differentiated smooth muscle cells. Sodium dodecyl sulfate (NaDodSO4)-polyacrylamide gel electrophoresis was used to compare the proteins synthesized and secreted by monolayer and nodular cultures of smooth muscle cells. Although most proteins appeared to be similar, the nodular cultures contained a unique heparin binding protein of Mr = 38,000 (38kD protein) (Millis, A.J.T., Hoyle, M., Reich, E., and Mann, D.M., 1985, J. Biol. Chem., 260:3754-3761). The 38kD protein was glycosylated and its apparent molecular weight was shifted to Mr = 32,500 after synthesis in the presence of tunicamycin or digestion with endoglycosidase F. The production of 38kD protein by nodular cell cultures did not appear to result from the degradation of a high molecular weight precursor in nodular conditioned medium. Further, it was not detected in monolayer cell conditioned medium that had been incubated with nodular cells. Finally, its synthesis was not induced in monolayer cell cultures that had been labeled in nodular cell conditioned medium. The 38kD protein appears to be uniquely associated with nodular cultures of smooth muscle cells.
在体外,血管平滑肌细胞的高密度单层培养物可被诱导形成多细胞结节。结节细胞在形态上似乎是分化的平滑肌细胞。使用十二烷基硫酸钠(NaDodSO4)-聚丙烯酰胺凝胶电泳来比较平滑肌细胞单层培养物和结节培养物合成与分泌的蛋白质。尽管大多数蛋白质看起来相似,但结节培养物中含有一种独特的肝素结合蛋白,其分子量为38,000(38kD蛋白)(米利斯,A.J.T.,霍伊尔,M.,赖希,E.,以及曼,D.M.,1985年,《生物化学杂志》,260:3754 - 3761)。38kD蛋白是糖基化的,在衣霉素存在下合成或用内切糖苷酶F消化后,其表观分子量移至32,500。结节细胞培养物产生38kD蛋白似乎不是由结节条件培养基中高分子量前体的降解所致。此外,在与结节细胞一起孵育的单层细胞条件培养基中未检测到该蛋白。最后,在结节细胞条件培养基中标记过的单层细胞培养物中也未诱导出其合成。38kD蛋白似乎与平滑肌细胞的结节培养物独特相关。