Expression of recombinant hyaluronan synthase (HAS) isoforms in CHO cells reduces cell migration and cell surface CD44
Expression of recombinant hyaluronan synthase (HAS) isoforms in CHO cells reduces cell migration and cell surface CD44
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DOI：
10.1006/excr.1999.4645
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发表时间：
1999-11-01
影响因子：
3.7
通讯作者：
Heldin, P
中科院分区：
文献类型：
--
作者：
Brinck, J;Heldin, P
在本研究中，我们研究了三种重组透明质酸脱氢酶（RAS蛋白）HAS 1，HAS 2和HAS 3的功能特性。HAS 3转染的CRO克隆表现出最高的透明质酸聚合速率，其次是HAS 2转染子，其比HAS 1转染子具有更高的催化活性。在活细胞中，所有三种RAS蛋白质都合成高分子量的透明质酸链（大于3.9 × 10（6））。在体外，HAS 2同种型产生分子量大于3.9 x 10（6）的透明质酸链，而HAS 3产生多分散透明质酸（分子量0.12-1 x 10（6）），HAS 1合成平均分子量为0.12 x 10（6）的短得多的链。因此，每种HAS蛋白可以与不同的细胞质蛋白相互作用，这可能影响它们的催化活性。具有合成约1 μ g透明质酸/1 × 10（5）细胞/24 h能力的CNO转染子被含透明质酸的包被包围，而产生约1/1低量透明质酸的转染子仅在硫酸软骨素蛋白聚糖存在下形成包被。透明质酸的生产一方面和细胞迁移和细胞表面CD 44的表达，另一方面之间的负相关性被发现; 4倍低迁移和2倍减少细胞表面CD 44受体被视为透明质酸生产增加1000倍以上的水平在未转染的细胞。透明质酸的产生和迁移与细胞的CD 44表达之间的反向关系对于调节细胞-细胞外基质相互作用是重要的。(C)北京：科学出版社.
In the present study we investigated the functional properties of the three recombinant hyaluronan synthases (RAS proteins) HAS1, HAS2, and HAS3. HAS3-transfected CRO clones exhibited the highest hyaluronan polymerization rate followed by HAS2 transfectants which were more catalytically active than HAS1 transfectants. In living cells all three RAS proteins synthesized hyaluronan chains of high molecular weight (larger than 3.9 x 10(6)). In vitro, the HAS2 isoform produced hyaluronan chains of a molecular weight larger than 3.9 x 10(6), whereas HAS3 produced polydisperse hyaluronan (molecular weight 0.12-1 x 10(6)), and HAS1 synthesized much shorter chains of an average molecular weight of 0.12 x 10(6). Thus, each HAS protein may interact with different cytoplasmic proteins which may influence their catalytic activity. CNO transfectants with the ability to synthesize about 1 mu g hyaluronan/1 x 10(5) cells/24 h were surrounded by hyaluronan-containing coats, whereas transfectants generating about Li-fold lower amounts of hyaluronan formed coats only in the presence of chondroitin sulfate proteoglycan. An inverse correlation between hyaluronan production on the one hand and cell migration and cell surface CD44 expression on the other was found; a 4-fold lower migration and a 2-fold decrease of cell surface CD44 receptors was seen when hyaluronan production increased 1000-fold over the level in the untransfected cells. The inverse relationships between hyaluronan production and migration and CD44 expression of cells are of importance for the regulation of cell-extracellular matrix interactions. (C) 1999 Academic Press.