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T A Beyer

Publications and source records attributed to T A Beyer.

22 records · Page 2Linked to original sources
PubMed1979

Biosynthesis of mammalian glycoproteins. Glycosylation pathways in the synthesis of the nonreducing terminal sequences.

Six purified glycosyltransferase (a beta-galactoside alpha 2 leads to 6 sialyltransferase, a beta-galactoside alpha 2 leads to 3 sialyltransferase, an alpha-N-acetylgalactosaminide alpha 2 leads to 6 sialyltransferase, a beta-galactoside alpha 1 leads to 2 fucosyltransferase, a beta-N-acetylglucosaminide alpha 1 leads to 3 fucosyltransferase, and a (fucosyl alpha 1 leads to 2) galactoside alpha 1 leads to 3 N-acetyl-galactosaminyltransferase) have been used to study the biosynthetic pathways for formation of the nonreducing terminal oligosaccharide sequences in mammalian glycoproteins. The two glycoproteins used as model acceptor substrates in this study were human asialotransferrin, which contains the nonreducing terminal oligosaccharide sequence Gal beta 1 leads to 4GlcNAc beta 1 leads to 2Man, and antifreeze glycoprotein, which contains oligosaccarides with the structure, Gal beta 1 leads to 3GalNAc alph 1 leads O-Thr. Sequential action of the six glycosyltransferases on these model substrates led to the formation of previously described oligosaccharide structures. The studies reported here indicate that the substrate specificities of the individual enzymes dictate the structures that can be synthesized and the pathways by which they may be formed. The actions of a number of the transferasesare mutually exclusive, thereby prohibiting the formation of theoretically possible oligosaccharide structures. Oligosaccharides with the terminal sequence NeuAc alpha 2 leads to 3(Fuc alpha 1 leads to 2)Gal beta 1 leads to 3GalNAc and NeuAc alpha 2 leads to 6Gal beta 1 leads to 4(Fuc alpha 1 leads to 3)GlcNAc cannot be formed because the prior incorporation of sialic acid by the sialyltransferases yields products that are not acceptor substrates for the fucosyltransferases, and vice versa. Synthesis of other products requires that the enzymes act sequentially in a specific order. The structures NeuAc alpha 2 leads to 6(Fuc alpha 1 leads to 2)Gal beta 1 leads to 4GlcNAc, Fuc alpha 1 leads to 2Gal beta 1 leads to 4(Fuc alpha 1 leads to 3)GlcNAc, GalNAc alpha 1 leads to 3(Fuc alpha 1 leads to 2)Gal beta 1 leads to 4GlcNAc, and GalNAc alpha 1 leads to 3(Fuc alpha 1 leads to 2)Gal beta 1 leads to 3GalNAc can only be synthesized if the fucosyl alpha 1 leads to 2 galactose linkage is formed first. Synthesis of the pentasaccharide sequences GalNAc alpha 1 leads to 3(Fuc alpha 1 leads to 2)Gal beta 1 leads to 3(NeuAc alpha 2 leads to 6)GalNAc and GalNAc alpha 1 leads to 3(Fuc alpha 1 leads to 2)Gal beta 1 leads to 4(Fuc alpha 1 leads to 3)GlcNAc requires that the N-acetylgalactosaminyltransferase act last on the former structure and that the alpha 1 leads to 3 fucosyltransferase act last on the latter. In those instances where a product can be formed by one of two possible pathways, the comparisons of reaction rates indicate that one pathway is usually preferred...

Carbohydrate Conformation↗
PubMed1976

The kinetic mechansim of bovine milk galactosyltransferase. The role of alpha-lactalbumin.

Initial rate parameters obtained with bovine galactosyltransferase at saturating Mn2+ concentrations, and a variety of acceptors including N-acetylglucosamine, glucose, ovalbumin, and di-N-acetylglucosamine are inconsistent with an ordered addition of UDP-galactose and acceptor substrates to the enzyme-Mn2+ complex. Inhibition patterns with N-acetylglucosamine or UDP-glucose as inhibitors of the galactosylation of ovalbumin indicated that either UDP-galactose or N-acetylglucosamine can bind to an enzyme-Mn2+ complex by a random equilibrium mechanism. Initial rate studies also indicate that alpha-lactalbumin may bind to either an enzyme-Mn2+-acceptor complex or an enzyme-Mn2+-UDP-galactose complex, suggesting that lactose synthesis also proceeds by a random equilibrium addition of substrates and alpha-lactalbumin. From the initial rate data assuming the random equilibrium mechanism, the dissociation constants for UDP-galactose, acceptor substrates, and alpha-lactalbumin from the appropriate complexes have been calculated. These values are in good agreement with those obtained independently by nonkinetic methods, providing additional support for the proposed random equilibrium mechanism. From similar studies with a cross-linked complex of alpha-lactalbumin and transferase, dissociation constants for UDP-galactose and acceptor substrates from the enzyme-Mn2+-alpha-lactalbumin complex were calculated. Comparison of each of the dissociation constants in the substrate addition phase shows that the binding of acceptor substrates and alpha-lactalbumin to enzyme-Mn2+ complexes is highly synergistic; the affinity of alpha-lactalbumin for the enzyme-Mn2+ acceptor complex is about 2 orders of magnitude greater than for the enzyme-Mn2+ complex. Similarly, the affinity of the acceptor for the enzyme-Mn2+-alpha-lactalbumin complex is about 2 orders of magnitude greater than the enzyme-Mn2+ complex. Synergism is also observed between alpha-lactalbumin and UDP-galactose binding but the synergism is much less than that observed with acceptor substrates and alpha-lactalbumin. Thus, the large decrease in the Michaelis constant for glucose in the presence of alpha-lactalbumin, which is observed for lactose synthesis by the galactosyltranferase, is primarily the result of the high degree of synergism in the binding of alpha-lactalbumin and glucose to enzyme-Mn2+ complexes. This synergism also accounts for the activation of N-acetyllactosamine synthesis by alpha-lactalbumin at low concentrations (less than 2 mM) of N-acetylglucosamine. An abortive enzyme-Mn2+-UDP-acceptor complex in the product release phase of the reaction appears to account for the inhibition of either lactose, or N-acetyllactosamine synthesis at a high concentration of either N-acetylglucosamine or glucose. This abortive complex is further stabilized by alpha-lactalbumin, thus the resulting substrate inhibition is observed at much lower acceptor concentrations in the presence of alpha-lactalbumin.

Acetylglucosamine↗
PubMedDate not supplied

Acquired alteration in the expression of blood groups in a patient with sideroblastic anemia and chronic renal failure.

A patient with well-differentiated lymphocytic lymphoma in remission presented with renal failure, sideroblastic anemia, and an acquired diminution in the expression of A antigen on his red cells. Other red cell antigens (H, M, i, and I) also were expressed weakly. Lewis antigen was expressed only in secretions. The diminished expression of the A antigen could not be accounted for by diminution in the specific glycosyl transferases responsible for its expression, since the activity in the serum of both the H enzyme (alpha 1 leads to 2 fucosyltransferase) and the A enzyme (alpha 1 leads to 3 N-acetylgalactosaminyltransferase) was not diminished. When sialic acid was removed by enzymatic treatment, the expression of the A and i antigens was augmented markedly, suggesting that the sialic acid may have altered the expression or detection of the antigens. This abnormality was probably related to dysplasia of the bone marrow which was otherwise manifest by ringed sideroblasts, unsegmented megakaryocytes, and diminished growth in vitro.

ABO Blood-Group System↗