Tropoelastin interacts with cell-surface glycosaminoglycans via its COOH-terminal domain
Tropoelastin interacts with cell-surface glycosaminoglycans via its COOH-terminal domain
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DOI：
10.1074/jbc.m507309200
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发表时间：
2005-12-09
影响因子：
4.8
通讯作者：
Mecham, RP
中科院分区：
文献类型：
--
作者：
Broekelmann, TJ;Kozel, BA;Mecham, RP
利用生物化学和细胞生物学方法，我们在原弹性蛋白的羧基末端确定了一个细胞相互作用位点。细胞与羧基末端序列的相互作用不是通过弹性蛋白结合蛋白（EBP67）进行的，因为无论是类似VGVAPG的肽段还是半乳糖苷糖类都没有改变黏附情况。我们的研究结果还表明，整合素不会促进细胞对原弹性蛋白的黏附。通过使用糖胺聚糖生物合成有缺陷的中国仓鼠卵巢突变细胞系，以及竞争研究和对特定细胞表面糖胺聚糖的酶促去除，细胞表面上与原弹性蛋白结合的部分被确定为含硫酸乙酰肝素和硫酸软骨素的糖胺聚糖，其中硫酸乙酰肝素更受青睐。肝素亲和层析结合细胞黏附测定法确定了最后17个氨基酸是原弹性蛋白羧基末端负责黏附活性的序列元件。
Using a biochemical and cell biological approach, we have identified a cell interaction site at the carboxyl terminus of tropoelastin. Cell interactions with the COOH-terminal sequence are not through the elastin-binding protein (EBP67) because neither VGVAPG-like peptides nor galactoside sugars altered adhesion. Our results also show that cell adhesion to tropoelastin is not promoted by integrins. Through the use of mutant Chinese hamster ovary cell lines defective in glycosaminoglycan biosynthesis, as well as competition studies and enzymatic removal of specific cell-surface glycosaminoglycans, the tropoelastin-binding moieties on the cell surface were identified as heparan and chondroitin sulfate-containing glycosaminoglycans, with heparan sulfate being greatly preferred. Heparin affinity chromatography combined with cell adhesion assays identified the last 17 amino acids as the sequence element at the carboxyl terminus of tropoelastin responsible for the adhesive activity.