title: Expression of recombinant hyaluronan synthase (HAS) isoforms in CHO cells reduces cell migration and cell surface CD44.
abstract: In the present study we investigated the functional properties of the three recombinant hyaluronan synthases (HAS proteins) HAS1, HAS2, and HAS3. HAS3-transfected CHO clones exhibited the highest hyaluronan polymerization rate followed by HAS2 transfectants which were more catalytically active than HAS1 transfectants. In living cells all three HAS 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. CHO transfectants with the ability to synthesize about 1 microgram hyaluronan/1 x 10 (5) cells/24 h were surrounded by hyaluronan-containing coats, whereas transfectants generating about 4-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.
authors.affiliation: Department of Medical Biochemistry and Microbiology, Unit of Biochemistry, Uppsala University, Biomedical Centre, Uppsala, S-75123, Sweden.
authors.forename: J
authors.initials: J
authors.lastname: Brinck
authors.forename: P
authors.initials: P
authors.lastname: Heldin
mesh_terms: D000818:Animals; D016466:CHO Cells; D002465:Cell Movement / Q000235:genetics*; D006224:Cricetinae; D015971:Gene Expression Regulation, Enzymologic; D014453:Glucuronosyltransferase / Q000096:biosynthesis* / Q000235:genetics*; D016695:Glycosyltransferases; D018960:Hyaluronan Receptors / Q000096:biosynthesis / Q000235:genetics; D000076002:Hyaluronan Synthases; D008565:Membrane Proteins; D020033:Protein Isoforms / Q000096:biosynthesis / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D014166:Transferases; D029867:Xenopus Proteins
chemical_list: D018960:Hyaluronan Receptors; D008565:Membrane Proteins; D020033:Protein Isoforms; D011994:Recombinant Proteins; D029867:Xenopus Proteins; D014166:Transferases; D016695:Glycosyltransferases; D014453:Glucuronosyltransferase; C056944:HAS1 protein, Xenopus; D000076002:Hyaluronan Synthases