Nemec R E, Toole B P, Knudson W
Department of Biochemistry, Rush-Presbyterian, St. Luke's Medical Center, Chicago, Il 60612.
Biochem Biophys Res Commun. 1987 Nov 30;149(1):249-57. doi: 10.1016/0006-291x(87)91632-9.
High-affinity, cell surface binding sites for hyaluronate were demonstrated on highly invasive human bladder carcinoma cells. These binding sites were shown to be specific for hyaluronate, saturable and exhibit a Km of 0.94 x 10(-9) M and a Bmax of 65 ng hyaluronate/10(6) cells. The binding of [3H]hyaluronate to a fixed cell-affinity column was competed with unlabeled hyaluronate and hyaluronate-hexasaccharide but not with hyaluronate-tetrasaccharide, chondroitin sulfate, heparin or non-sulfated dextran. Pre-treatment of cells with protease destroyed the binding activity whereas pretreatment with Streptomyces hyaluronidase to reveal occupied binding sites had no effect. No hyaluronate-binding activity was observed on normal human fibroblasts.
在高侵袭性人膀胱癌细胞上证实存在透明质酸的高亲和力细胞表面结合位点。这些结合位点对透明质酸具有特异性,可饱和,其Km为0.94×10⁻⁹ M,Bmax为65 ng透明质酸/10⁶个细胞。[³H]透明质酸与固定化细胞亲和柱的结合可被未标记的透明质酸和透明质酸六糖竞争,但不能被透明质酸四糖、硫酸软骨素、肝素或非硫酸化葡聚糖竞争。用蛋白酶预处理细胞会破坏结合活性,而用链霉菌透明质酸酶预处理以揭示占据的结合位点则没有效果。在正常人成纤维细胞上未观察到透明质酸结合活性。