Mikuni-Takagaki Y, Toole B P
J Cell Biol. 1980 May;85(2):481-8. doi: 10.1083/jcb.85.2.481.
Hyaluronate is associated with the cell surface of cultured Rous sarcoma virus-transformed chondrocytes. Detachment of these cells from their substratum by a variety of reagents is accompanied by release of 75-100% of this hyaluronate into solution. Treatment of the cells with 200 U/ml protease-free Streptomyces hyaluronidase at 37 degrees C cause release of greater than 90% of the cell surface hyaluronate and complete cell detachment. Treatment with a lower concentration of Streptomyces hyaluronidase (30 U/ml) at 25 degrees C or a corresponding activity of testicular hyaluronidase gives similar results, but only in the presence of mM EGTA. Treatment with the lower activities of either hyaluronidase or with 1 mM EGTA alone release only approximately 45% of the cell surface hyaluronate and does not cause significant cell detachment. It is concluded that there are two populations of cell surface hyaluronate differing in their accessibility or their resistance to dissociation from other components of the cell surface. It is proposed that the less readily released fraction is located between the transformed chondrocyte surface and substratum and is necessary for their interaction.
透明质酸盐与培养的劳斯肉瘤病毒转化的软骨细胞的细胞表面相关。用多种试剂使这些细胞从其基质上脱离时,会伴随75%-100%的这种透明质酸盐释放到溶液中。在37℃用200 U/ml无蛋白酶的链霉菌透明质酸酶处理细胞,会导致超过90%的细胞表面透明质酸盐释放以及细胞完全脱离。在25℃用较低浓度的链霉菌透明质酸酶(30 U/ml)或相应活性的睾丸透明质酸酶处理,会得到相似结果,但仅在存在毫摩尔级EGTA的情况下。用较低活性的任何一种透明质酸酶或单独用1 mM EGTA处理,仅释放约45%的细胞表面透明质酸盐,且不会导致明显的细胞脱离。结论是存在两种细胞表面透明质酸盐群体,它们在可及性或与细胞表面其他成分解离的抗性方面存在差异。有人提出,较难释放的部分位于转化的软骨细胞表面与基质之间,是它们相互作用所必需的。