Study of the fucosyltransferase system in intestinal microsomes.
Fucosyltransferase activity of rat small intestine microsomes is solubilized by 0.5% Triton X-100. The solubilized activity can be purified up to 8,300-fold using DEAE-cellulose and affinity chromatography on GDP-Sepharose. At this step, chromatography on Sephadex G15 separates different specificities: N-acetylglucosaminide-alpha-(1,3)-fucosyltransferase acting on asialoserotransferrin, and galactoside-alpha-(1,2)-fucosyltransferase acting on O-glycans of asialofetuin. The use of small saccharidic acceptors also indicates the presence of a N-acetylglucosaminide-alpha-(1,4)-fucosyltransferase and of a very weak glucose-alpha-(1,3)-fucosyltransferase activity. These activities are tightly bound to concanavalin A-Sepharose, suggesting that they are supported by N-glycosylproteins.
Martin
A
A
Department of General and Medical Biochemistry, INSERM-CNRS U 189, Lyon-Sud Medical School, Oullins, France.
Biol
M C
MC
Broquet
P
P
Richard
M
M
Louisot
P
P
D000818:Animals; D002848:Chromatography, DEAE-Cellulose; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D005647:Fucosyltransferases / Q000302:isolation & purification / Q000378:metabolism*; D006602:Hexosyltransferases / Q000378:metabolism*; D007422:Intestines / Q000201:enzymology*; D007700:Kinetics; D008297:Male; D008861:Microsomes / Q000201:enzymology*; D051381:Rats; D011919:Rats, Inbred Strains; D013379:Substrate Specificity
D005647:Fucosyltransferases; D006602:Hexosyltransferases