Turley E A
Department of Pharmacology, Faculty of Medicine, University of Calgary, Alberta, Canada.
J Biol Chem. 1989 May 25;264(15):8951-5.
The addition of hyaluronate to intact chick embryonic heart fibroblasts enriched with a hyaluronate-binding protein (HABP) stimulated phosphorylation of tyrosine and serine/threonine residues in cellular proteins. A protein complex containing a hyaluronate-binding protein (cell-HABP) was isolated from the cultured heart fibroblasts. The isolated complex (Mr approximately 1 x 10(6] contained phosphoproteins that exhibited protein kinase activity specifically stimulated by hyaluronate. Both tyrosine and serine residues in the protein complex were phosphorylated in response to this glycosaminoglycan. The hyaluronate-stimulated protein kinase activity was tightly associated with cell-HABP in vitro; enzyme activity co-immunoprecipitated with cell-HABP using a monospecific anti-HABP antibody and co-eluted with cell-HABP when chromatographed on a column of Sephacryl S-1000 in 2.0 M guanidine hydrochloride. The uniqueness of the cell-HABP-associated protein kinase activity was suggested by both its specific response to hyaluronate, relative to related glycosaminoglycans such as heparin and chondroitin sulfate or to growth factors such as epidermal growth factor or insulin, and its antigenic distinction from other protein kinases such as growth factor receptors. These results point to a new mechanism by which glycosaminoglycans, such as hyaluronate, may modify cell behavior.
向富含透明质酸结合蛋白(HABP)的完整鸡胚心脏成纤维细胞中添加透明质酸,可刺激细胞蛋白质中酪氨酸和丝氨酸/苏氨酸残基的磷酸化。从培养的心脏成纤维细胞中分离出一种含有透明质酸结合蛋白的蛋白质复合物(细胞-HABP)。分离出的复合物(分子量约为1×10⁶)含有磷蛋白,该磷蛋白表现出受透明质酸特异性刺激的蛋白激酶活性。蛋白质复合物中的酪氨酸和丝氨酸残基均因这种糖胺聚糖而发生磷酸化。在体外,透明质酸刺激的蛋白激酶活性与细胞-HABP紧密相关;使用单特异性抗HABP抗体时,酶活性与细胞-HABP共免疫沉淀,并且在2.0M盐酸胍中于Sephacryl S-1000柱上进行层析时,酶活性与细胞-HABP共洗脱。细胞-HABP相关的蛋白激酶活性的独特性体现在,相对于相关糖胺聚糖(如肝素和硫酸软骨素)或生长因子(如表皮生长因子或胰岛素),它对透明质酸有特异性反应,并且在抗原性上与其他蛋白激酶(如生长因子受体)不同。这些结果指出了一种新的机制,通过该机制,诸如透明质酸之类的糖胺聚糖可能会改变细胞行为。