Two alpha-3-D-galactosyltransferases in rabbit stomach mucosa with different acceptor substrate specificities.
Homogenates of rabbit stomach mucosa were examined for enzymes catalysing the transfer of D-galactose from UDP-D-galactose to various low-molecular-weight acceptors of known structure. Treatment of the products with alpha and beta-D-galactosidases revealed that D-galactose was transferred in both alpha and beta-anomeric linkages. The beta-D-galactosyltransferase used N-acetylglucosamine and compounds containing terminal nonreducing beta-N-acetylglucosaminyl residues as acceptor substrates. The compounds accepting D-galactose in alpha-anomeric linkage had unsubstituted terminal non-reducing beta-D-galactosyl units or a fucose substituent on the carbon-2 position of a subterminal beta-D-galactosyl unit. Methylation analysis of the products formed with N-acetyllactosamine [beta-D-Galp(1 leads to 4)D-GlcNAcp] and 2'fucosyllactose [alpha-L-Fucp(1 leads to 2)-beta-D-Galp(1 leads to 4)D-Glcp] revealed that D-galactose was transferred to the carbon-3 position of the beta-D-galactosyl residue in both of these acceptor substrates. Competition experiments with the two substrates indicated that the transfer of D-galactose was catalysed in each case by a different alpha-3-D-galactosyltransferase. Differences were also observed in the solubility properties of the enzymes: the alpha-3-D-galactosyltransferase using acceptor substrates with unsubstituted beta-D-galactosyl residues was more readily soluble both in the presence and absence of detergents than the transferase using beta-D-galactosyl residues substituted at carbon-2 with L-fucose. These findings demonstrate that rabbit stomach mucosa has two distinct alpha-3-D-galactosyltransferases: one, which is more tightly membrane-bound, resembles the human B-gene-specified transferase in its acceptor specificity, and the second, which is a more soluble enzyme, transfers D-galactose to the same positional linkage in unsubstituted beta-D-galactosyl residues.
D000818:Animals; D002499:Centrifugation, Density Gradient; D005700:Galactosyltransferases / Q000145:classification / Q000302:isolation & purification* / Q000378:metabolism; D005753:Gastric Mucosa / Q000201:enzymology*; D011817:Rabbits; D012995:Solubility; D013379:Substrate Specificity
D005700:Galactosyltransferases; C023155:blood-group-substance alpha-D-galactosyltransferase; C037959:beta-D-galactosyl-N-acetylglucosaminylglycopeptide alpha-1,3-galactosyltransferase