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Differential processing and regulation of thyroid-stimulating hormone subunit carbohydrate chains in thyrotropic tumors and in normal and hypothyroid pituitaries.

Abstract
Thyroid-stimulating hormone (TSH) alpha- and beta-subunit glycosylation was investigated in mouse thyrotropic tumor and in normal and hypothyroid pituitary cells for various periods of time in the presence of [3H]mannose or [3H]galactose. After sequential precipitation with anti-alpha and anti-beta sera, subunits were treated with Pronase followed by endo-beta-N-acetylglucosaminidase H (Endo H) and analyzed by paper chromatography. In primary cultures of thyrotropic tumor cells incubated for 60 min with [3H]mannose, primarily Man9GlcNAc and Man8GlcNAc were found on TSH + alpha subunits, whereas Glc1Man9GlcNAc and Man9GlcNAc were prominent on free beta subunits. After preincubation of cells for 16 h in the presence or absence of glucose followed by a 60-min pulse of [3H]mannose, there was an 8-fold increase in labeled TSH + alpha but only a minimal change in free beta or total proteins. In the absence of glucose, there was a selective accumulation of Man8GlcNAc on TSH + alpha but not on free beta or total proteins; however, there was no detectable accumulation of Endo H resistant forms during glucose starvation on TSH subunits or total proteins. Normal mouse and rat pituitary minces incubated for 60 min with either [3H]mannose or [3H]galactose showed no glucose-containing species on TSH subunits, but equal amounts of Man9GlcNAc and Man8GlcNAc on TSH + alpha, and mostly Man9GlcNAc on free beta subunits. In contrast, hypothyroid mouse and rat pituitaries exhibited an increase in Glc1Man9NAc and Glc1Man8GlcNAc on free beta but not on TSH + alpha or total proteins.(ABSTRACT TRUNCATED AT 250 WORDS)
AuthorsC Ronin, B S Stannard, B D Weintraub
JournalBiochemistry (Biochemistry) Vol. 24 Issue 20 Pg. 5626-31 (Sep 24 1985) ISSN: 0006-2960 [Print] United States
PMID4074717 (Publication Type: Journal Article)
Chemical References
  • Macromolecular Substances
  • Oligosaccharides
  • Tritium
  • Thyrotropin
  • Mannose
  • Galactose
Topics
  • Animals
  • Female
  • Galactose (metabolism)
  • Hypothyroidism (metabolism)
  • Macromolecular Substances
  • Male
  • Mannose (metabolism)
  • Mice
  • Mice, Inbred Strains
  • Oligosaccharides (biosynthesis)
  • Pituitary Gland (metabolism)
  • Pituitary Neoplasms (metabolism)
  • Protein Processing, Post-Translational
  • Rats
  • Rats, Inbred Strains
  • Reference Values
  • Thyrotropin (genetics)
  • Tritium

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