Differential effect of inhibitors of oligosaccharide processing on the secretion of thyrotropin from dispersed rodent pituitary cells.
We examined the effect of various inhibitors of oligosaccharide processing on the content and secretion of newly synthesized thyroid-stimulating hormone (TSH) from dispersed hypothyroid rodent pituitary cells. 1-deoxynojirimycin and N-methyl-1-deoxynojirimycin, both inhibitors of glucosidases I and II, decreased intracellular TSH (to 60-76% of control) and secreted TSH (to 60-63% of control) after a 1-hour incubation (pulse) with [35S]methionine and an 8-hour incubation (chase) in isotope-free media. In contrast, deoxymannojirimycin and swainsonine, inhibitors of mannosidase I and II, respectively, increased both intracellular TSH (to 267-309% of control) and secreted TSH (to 192% of control) at 8 hours. TSH oligosaccharides synthesized in the presence of these glucosidase and mannosidase inhibitors were largely sensitive to endo-beta-N-acetylglucosaminidase H (endo H), confirming inhibition of processing. Despite differences in oligosaccharide structure, the in vitro bioactivities of these secreted TSH isoforms were nearly identical. These data confirm and extend previous work performed with 1-deoxynojirimycin suggesting that glucosylated high mannose forms of TSH are more susceptible to intracellular degradation. The novel finding that deoxymannojirimycin and swainsonine increase secreted and total TSH above control levels suggests that non-glucosylated high mannose forms as well as hybrid-type oligosaccharides may facilitate secretion and direct TSH away from a natural degradation pathway.
Stannard
B S
BS
Molecular, Cellular and Nutritional Endocrinology Branch, National Institute of Diabetes and Digestive and Kidney Diseases, Bethesda, MD 20892.
Gesundheit
N
N
Thotakura
N R
NR
Gyves
P W
PW
Ronin
C
C
Weintraub
B D
BD
D017485:1-Deoxynojirimycin / Q000031:analogs & derivatives; D000470:Alkaloids / Q000494:pharmacology; D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D005944:Glucosamine / Q000031:analogs & derivatives / Q000494:pharmacology; D006026:Glycoside Hydrolases / Q000037:antagonists & inhibitors; D006031:Glycosylation; D007037:Hypothyroidism / Q000378:metabolism; D008297:Male; D008361:Mannosidases / Q000037:antagonists & inhibitors; D051379:Mice; D009844:Oligosaccharides / Q000378:metabolism*; D010902:Pituitary Gland / Q000187:drug effects / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D051381:Rats; D011919:Rats, Inbred Strains; D017026:Swainsonine; D013964:Thyroid Neoplasms / Q000378:metabolism*; D013965:Thyroidectomy; D013972:Thyrotropin / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism
D000470:Alkaloids; D000900:Anti-Bacterial Agents; D009844:Oligosaccharides; D017485:1-Deoxynojirimycin; D013972:Thyrotropin; D006026:Glycoside Hydrolases; D008361:Mannosidases; D005944:Glucosamine; D017026:Swainsonine; C033424:N-methyldeoxynojirimycin