Search

A cell line, IGROV1, originating from a human ovarian cancer, releases a galactosyltransferase activity in its culture medium during proliferation. The proliferating IGROV1 cells appear as two populations: some cells grow in floating clusters whereas the greater part of them adhere to the culture substrate. The study of galactose transfer by intact cells onto an exogenous glycoprotein acceptor (ovomucoid) shows the presence of surface-associated galactosyltransferase onto the two cellular sub-populations. Opposite to intracellular activity, surface-associated and released galactosyltransferase activities depend on cellular adhesion and proliferation.

A soluble galactosyltransferase exists in the cell sap of the alveolar cells of the lung. The optimal conditions of galactosyltransferase are: pH 6, 20 DEGREES C, 10 mM Mn2+, 1 mM Mg2+. Michaelis constant for substrate UDP-galactose is 390 nM. By electrofocusing on Ampholine column, galactosyltransferase appears in a unique peak, at pHi 4.8. The parameters of the purified enzyme are the same as that of the crude enzyme. UDP is a noncompetitive inhibitor for the purified enzyme.

The authors have selected three series of families presenting several heterozygous AB gene combinations: the same gene A and the same gene B; the same gene B and different genes A; and the same gene A and different genes B. The results concerning these families show that the quantitiy of A1 and A substances (for A1B) or A (for A2B) is related to the "quality" of the alpha-D-galactosyl transferase acting on a more or less important H substratum. The alpha-N-acetyl-D-galactosaminyl transferase seems to have a minor function.

Galactosyltransferase which catalyzes the transfer from UDP-galactose to either endogeneous glycoproteins, free N-acetylglucosamine or N-acetylglucosaminyl residues in the carbohydrate portion of glycoproteins, or to glucose when alpha-lactalbumin is added, occurs in human milk fat globule membranes. Various treatments (washing of membranes, freezing and thawing) did not affect this activity. In the presence of Triton X-100, the enzyme shows appreciable latency, This detergent was then used to solubilize the enzyme and to study its main characteristics. A competition and a heat stability experiment show that only one enzyme acts on two substrates (free N-acetylglucosamine or desialyzed and degalactosylated fetuin). UDP-galactose hydrolase activities were very low compared to those of the bovine milk fat globule membranes. Other characteristic enzymes of Golgi vesicles were found in human milk fat globules membranes. It is of interest to find out whether this is the result of contamination with cytoplasmic particles or whether it reflects the participation of Golgi vesicles in human milk fat globule secretion.

Cell cultures of foetal rabbit aorta are cultivated with a lathyric agent (beta-amino-propio-nitrile) or with an hypercholesterolemic serum; if morphological features, in these two cases, correspond with modifications observed, in vivo, when adult rabbits are respectively submitted to the same treatment, enzymatic activities of collagen metabolism vary in opposite way. Therefore, the influence of different parameters to be studied on vascular cell functions become easier.

Tranfer of (14C)-gal from exogenous UDP(14C)-gal has been demonstrated with ovomucoid as glycoprotein acceptor in ascitic fluid sera from Balb/c mice bearing isogenic YC8 tumor. Transfer exists without ovomucoid, in great ratio in ascitic fluid and in sera from ascitic mice too, showing occurrence of endogenous acceptors, the origin of which has to be proved.

The existence of UDP-galactose:galactosyltransferase activities at the external surface of cultured neurons from chick embryo hemispheres, was demonstrated. [14C]-galactose was transferred upon cellular substrates present on the neuronal surface and upon exogenous substrates (ovomucoïd) bound to Sepharose beads. The activity of ectogalactosyltransferase was not due to a leakage of intracellular enzymes into the incubation medium nor to an intracellular enzymatic activity utilizing [14C]-galactose obtained from the hydrolysis of UDP-[14C]-galactose. Kinetics of these ectogalactosyltransferases were analysed. The results support the hypothesis of a communication and/or interaction among the nerve cell surfaces via glycoconjugates or their biosynthetic enzymes.

We have demonstrated, by the transfer of (1-14C) GalN Ac from exogenous UDP[ (1-14C) GalN Ac into a glycopeptide H-acceptor, the presence of an alpha-N-acetylgalactosaminyltransferase in the wall and fluid of ovarian tumors. The transfer occurs in the absence of exogenous acceptor but to a greater degree in the tumor wall and fluid obtained from blood group O patients, thereby indicating the presence of preferential endogenous acceptor(s) in tumors of these subjects.