In the proposed program we plan to study novel human glycosyltransferases. Much emphasis will be made on the identification and characterization of enzymes present in ovarian epithelial linings and fluids. Synthesis of various oligosaccharides which can be effectively employed as acceptor-substrates for certain glycosyltransferases have already been accomplished in our laboratory. A number of these synthetic substrates have proven to be very useful and valuable acceptors for glycosyltransferases from sources other than human. Our main strategy for the characterization of the product resulting from the reaction of a glycosyltransferase and its acceptor involves the synthesis of the expected saccharide which can be used as a reference compound. The structure of the saccharide isolated from enzymatic product will be established by n.m.r., mass spectroscopy, and methylation studies. We plan to employ glycosidases for the structural studies of the new product. Adequate use of newly acquired HPLC instrumentation will be made to facilitate these investigations. The proposed studies are directed toward the characterization of the spectrum of glycosyltransfeerases which are responsible for elongation of carbohydrate units O-glycosidically linked to Serine or Threonine. We are concentrating on the study of glycosyltransferases which have not been identified but whose existence is inferred from structural studies and the """"""""one enzyme-one linkage"""""""" concept. Our studies will further elucidate the biosynthetic pathway of glycoconjugates. It is anticipated that glycosyltransferases of defined specificity, as proposed in this program in conjunction with glycosidases, may be of general value in further understanding the roles of specific oligosaccharide structures in the complex mileu of cell-surface glycoconjugates.
Yazawa, S; Abbas, S A; Madiyalakan, R et al. (1986) N-acetyl-beta-D-glucosaminyltransferases related to the synthesis of mucin-type glycoproteins in human ovarian tissue. Carbohydr Res 149:241-52 |
Brockhausen, I; Matta, K L; Orr, J et al. (1986) Mucin synthesis. Conversion of R1-beta 1-3Gal-R2 to R1-beta 1-3(GlcNAc beta 1-6)Gal-R2 and of R1-beta 1-3GalNAc-R2 to R1-beta 1-3(GlcNAc beta 1-6)GalNAc-R2 by a beta 6-N-acetylglucosaminyltransferase in pig gastric mucosa. Eur J Biochem 157:463-74 |
Brockhausen, I; Matta, K L; Orr, J et al. (1985) Mucin synthesis. UDP-GlcNAc:GalNAc-R beta 3-N-acetylglucosaminyltransferase and UDP-GlcNAc:GlcNAc beta 1-3GalNAc-R (GlcNAc to GalNAc) beta 6-N-acetylglucosaminyltransferase from pig and rat colon mucosa. Biochemistry 24:1866-74 |