Studies on hormonal modulation of asparagine-linked glycoprotein biosynthesis in explant cultures of rat mammary gland.
Glucosidase I has been purified to homogeneity and polyclonal antibodies against the enzyme have been prepared. The anti-glucosidase I antibodies recognized a single band of 85 kDa on western blot at a dilution as high as 1:2000 and also inhibited the enzyme activity, suggesting the specificity of the antibodies. Con A-Sepharose binding experiment indicates that this enzyme itself is a high mannose type N-linked glycoprotein. The increase in the electrophoretic mobility of 85 kDa band following digestion with endoglycosidase H and F strengthened this observation. The presence of any O-linked sugar attached covalently to glucosidase I could not be detected by binding assays with O-linkage specific biotinylated lectins. The studies on developmental regulation suggest that the synthesis of glucosidase I is modulated with the ontogeny of the gland. Lactogenic hormones, viz. insulin, hydrocortisone and prolactin, appeared to regulate the synthesis of glucosidase I. The possible role of these hormones in the overall regulation of protein N-glycosylation has been discussed.
Shailubhai
K
K
Department of Animal Sciences, University of Maryland, College Park 20742.
Saxena
E S
ES
Balapure
A K
AK
Vijay
I K
IK
D000818:Animals; D001216:Asparagine / Q000378:metabolism; D015153:Blotting, Western; D046508:Culture Techniques; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006023:Glycoproteins / Q000096:biosynthesis*; D006854:Hydrocortisone / Q000494:pharmacology*; D007328:Insulin / Q000494:pharmacology*; D008321:Mammary Glands, Animal / Q000187:drug effects / Q000201:enzymology* / Q000254:growth & development; D011247:Pregnancy; D011388:Prolactin / Q000494:pharmacology*; D051381:Rats; D000520:alpha-Glucosidases / Q000187:drug effects / Q000378:metabolism*
D006023:Glycoproteins; D007328:Insulin; D001216:Asparagine; D011388:Prolactin; C042059:glucosidase I; D000520:alpha-Glucosidases; D006854:Hydrocortisone