Three different endogenous alpha-L-fucosyltransferases expressed in COS cells.
The monkey kidney COS cell line is frequently used for the transient expression of cloned human fucosyltransferase cDNAs in the belief that negligible endogenous expression of fucosyltransferase genes occurs in these cells. In the course of transfection experiments we observed weak cell surface expression of sialyl-Lex and weak fucosyltransferase activity in extracts of control untransfected cells. Since these activities could complicate interpretation of the results with the transfected genes, a more detailed examination was undertaken that has now revealed expression of three different fucosyltransferases in the cells. One enzyme, which utilises N-acetyllactosamine as substrate, has a pH optimum of 7.0, is resistant to heat inactivation, and has been tentatively identified as an alpha1,3-fucosyltransferase. A second enzyme which acts on asialo-fetuin has a pH optimum of 5.5 and is rapidly inactivated by heat; the acceptor sugar and positional linkage of the transferred fucose are not yet established. A third enzyme that utilises asialo-agalacto-fetuin as acceptor is provisionally identified as an alpha1,6-fucosyltransferase.
Clarke
J L
JL
Royal Postgraduate Medical School, Hammersmith Hospital, London, W12 ONN, United Kingdom.
Watkins
W M
WM
D000818:Animals; D019556:COS Cells; D002457:Cell Extracts; D005033:Ethylmaleimide / Q000494:pharmacology; D005434:Flow Cytometry; D005647:Fucosyltransferases / Q000276:immunology / Q000378:metabolism*; D006358:Hot Temperature; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007527:Isoenzymes / Q000276:immunology / Q000378:metabolism*; D009844:Oligosaccharides / Q000378:metabolism; D000080506:Sialyl Lewis X Antigen
D002457:Cell Extracts; D007527:Isoenzymes; D009844:Oligosaccharides; D000080506:Sialyl Lewis X Antigen; D005647:Fucosyltransferases; D005033:Ethylmaleimide