Identification and characterization of a Drosophila melanogaster ortholog of human beta1,4-galactosyltransferase VII.
Drosophila melanogaster is widely considered to be an attractive model organism for studying the functions of the carbohydrate moieties of glycoconjugates produced by higher eukaryotes. However, the pathways of glycoconjugate biosynthesis are not as well defined in insects as they are in higher eukaryotes. One way to address this problem is to identify genes in the Drosophila genome that might encode relevant functions, express them, and determine the functions of the gene products by direct biochemical assays. In this study, we used this approach to identify a putative Drosophila beta4-galactosyltransferase gene and determine the enzymatic activity of its product. Biochemical assays demonstrated that this gene product could transfer galactose from UDP-galactose to a beta-xylosyl acceptor, but not to other acceptors in vitro. The apparent K(m) values for the donor and acceptor substrates indicated that this gene product is a functional galactosyltransferase. Additional assays showed that the enzyme is activated by manganese, has a slightly acidic pH optimum, and is localized in the insect cell Golgi apparatus. These results showed that Drosophila encodes an ortholog of human beta4-galactosyltransferase-VII, also known as galactosyltransferase I, which participates in proteoglycan biosynthesis by transferring the first galactose to xylose in the linkage tetrasaccharide of glycosaminoglycan side chains.
Vadaie
Nadia
N
Department of Molecular Biology, University of Wyoming, P.O. Box 3944, Laramie, WY 82071-3944, USA.
Hulinsky
Rebecca S
RS
Jarvis
Donald L
DL
D000595:Amino Acid Sequence; D000818:Animals; D029721:Drosophila Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D004331:Drosophila melanogaster / Q000201:enzymology*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D009237:N-Acetyllactosamine Synthase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D010802:Phylogeny; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity
D029721:Drosophila Proteins; C473070:beta4GalT7 protein, Drosophila; D009237:N-Acetyllactosamine Synthase