29 research outputs found
The carbohydrate moiety of bovine thyrotropin is essential for full bioactivity but not for receptor recognition.
No full textTSH is a glycoprotein hormone whose carbohydrate content varies among different species. Although recent studies suggest that variants of TSH deficient in carbohydrate occur naturally, the significance of the carbohydrate moiety of TSH in respect to its thyrotropic function is unclear. The present studies were undertaken, therefore, to examine this question. A highly purified preparation of bovine TSH (bTSH) was deglycosylated by treatment with anhydrous hydrogen fluoride. Amino acid and carbohydrate analyses of the original and deglycosylated preparations indicated that approximately 85\% of the carbohydrate originally present had been removed and that the protein moiety was unaltered. As judged from TSH radioreceptor assays, bTSH and deglycosylated bTSH (dg-bTSH) bound to human thyroid membranes with equal affinity, since both caused a half-maximal inhibition of [125I]bTSH binding at approximately equal concentrations. Nonetheless, dg-bTSH at optimal concentration displayed only about one third the activity of intact TSH in stimulating adenylate cyclase activity in human thyroid membranes. dg-bTSH also antagonized the adenylate cyclase-stimulating activity of intact bTSH in this system, but only weakly, since abolition of the bTSH effect required an approximately 40-fold higher concentration of dg-bTSH. In cultures of FRTL5 cells, a cloned line of follicular cells derived from normal rat thyroid, both intact and dg-bTSH enhanced cell growth, as measured by [3H]thymidine incorporation and stimulated cAMP release in the medium, but the response elicited by dg-bTSH was much less than that caused by equal concentrations of the intact hormone. In accord with the findings in the in vitro assays, dg-bTSH evoked a much smaller response than bTSH did in the in vivo mouse assay. It is concluded that although not required for receptor recognition, the carbohydrate moiety of bTSH is essential for the full expression of its biological activity
Circadian properties of vasopressin and melatonin rhythms in cat cerebrospinal fluid
No full text Using a method for continuous removal of cisternal cerebrospinal fluid (CSF) from freely moving cats, we delineated the circadian nature of the daily rhythm in CSF arginine vasopressin. The daily melatonin rhythm was also monitored in CSF as another marker of circadian function. Under diurnal lighting conditions, both hormones exhibited prominent daily rhythms; the CSF vasopressin rhythm was characterized by high daytime values, whereas the CSF melatonin rhythm was characterized by high nighttime levels. In contrast, drinking behavior exhibited a 24-h component in only one of four animals studied. Daily CSF rhythms of vasopressin and melatonin persisted for over 78 days of study in constant light. The vasopressin rhythm clearly free-ran in this environment, manifesting cycle lengths of slightly greater than 24 h. The daily melatonin pattern split into several components with increasing time in constant light. An acute 8-h phase delay in the daily light-dark cycle resulted in corresponding but gradual phase shifts in both rhythms. These results indicate that both the vasopressin and melatonin rhythms in cat CSF are endogenously generated and are entrained by the daily light-dark cycle. </jats:p
A multiresponse parathyroid hormone assay: an inhibitor has agonist properties in vivo
No full textVitamin D-deficient rats subjected to thyroparathyroidectomy (TPTX) were used to evaluate in vivo the biological properties of native bovine parathyroid hormone (bPTH) and chemically synthesized fragments and analogues of the hormone on several parameters of hormone action: calcium and phosphorus fluxes, generation of cyclic adenosine 3',5'-monophosphate (cAMP), and the metabolism of 25-hydroxyvitamin D3 [25(OH)D3]. Vitamin D-deficient rats, after TPTX or sham operation, were intravenously infused with a nutrient containing 7.5 mM CaCl2 for 30 h. During the last 7 h, PTH or one of its analogues was infused intravenously at rates between 0.04 and 20 nmol/h. One hour after the start of the peptide infusion, tritiated 25(OH)D3 was injected. Urine was collected hourly for phosphate and cAMP determinations and, at the end of the experiment, blood was obtained to determine the relative accumulation of tritiated 1,25-dihydroxyvitamin D3 ([3H]1,25(OH)2D3). Infusion of bPTH-(1--84), bPTH-(1--34), human (h)PTH-(1--34), or [Nle8, Nle18, Tyr34]bPTH-(1--34) amide was accompanied by a comparable dose-dependent decrease in plasma phosphate and a dose-dependent increase in plasma calcium and [3H]-1,25(OH)2D3, and urinary excretion of phosphate and cAMP. An evaluation of [Nle8, Nle18, Tyr34]bPTH-(3--34) amide, a potent inhibitor of PTH action in vitro in the renal adenylate cyclase assay, revealed that the analogue possessed weak agonist properties in vivo. The analogue increased excretion of both cAMP and phosphate in the urine, decreased plasma phosphate levels, and increased the accumulation of [3H]-1,25(OH)2D3 in the plasma. This multiparameter model system should aid in the elucidation of the in vivo biological effects of PTH and its analogues.</jats:p
Role of parathyroid hormone in phosphate transport across rat duodenum
No full text The effect of parathyroid extract (PTE) on phosphate transport was studied by perfusing everted duodenal loops of rats in vitro. It was found that PTE had no effect on the membrane potential or on the pH of the medium. However, PTE increased the influx (transfer from mucosa to serosa) 70%, the uptake by the tissue from the mucosal side 30%, and the lactate transfer into the serosal fluid 30%. Outflux of phosphate (transfer from serosa to mucosa) was as great as the influx but unaffected by PTE. Phosphate uptake from the serosal side was a third of the mucosal uptake and also unaffected by PTE. Metabolic inhibitors, iodoacetate, arsenite, and DNP strongly inhibited phosphate influx and tissue uptake. Lactate formation and transfer were also inhibited by iodoacetate and arsenite but not by DNP. In addition the three inhibitors suppressed completely the PTE effects. It is suggested that phosphate movements across the intestine follow different pathways whose dependence upon metabolism, hormonal action, or electrochemical gradient might be different and independent. </jats:p
Alternative splicing and endoproteolytic processing generate tissue-specific forms of pituitary peptidylglycine alpha-amidating monooxygenase (PAM).
No full textInternational audienceThe pituitary is a rich source of peptidylglycine alpha-amidating monooxygenase (PAM). This bifunctional protein contains peptidylglycine alpha-hydroxylating monooxygenase (PHM) and peptidyl-alpha-hydroxyglycine alpha-amidating lyase (PAL) catalytic domains necessary for the two-step formation of alpha-amidated peptides from their peptidylglycine precursors. In addition to the four forms of PAM mRNA identified previously, three novel forms of PAM mRNA were identified by examining anterior and neurointermediate pituitary cDNA libraries. None of the PAM cDNAs found in pituitary cDNA libraries contained exon A, the 315-nucleotide (nt) segment situated between the PHM and PAL domains and present in rPAM-1 but absent from rPAM-2. Although mRNAs of the rPAM-3a and -3b type encode bifunctional PAM precursors, the proteins differ significantly. rPAM-3b lacks a 54-nt segment encoding an 18-amino acid peptide predicted to occur in the cytoplasmic domain of this integral membrane protein; rPAM-3a lacks a 204-nt segment including the transmembrane domain and encodes a soluble protein. rPAM-5 is identical to rPAM-1 through nt 1217 in the PHM domain; alternative splicing generates a novel 3'-region encoding a COOH-terminal pentapeptide followed by 1.1 kb of 3'-untranslated region. The soluble rPAM-5 protein lacks PAL, transmembrane, and cytoplasmic domains. These three forms of PAM mRNA can be generated by alternative splicing. The major forms of PAM mRNA in both lobes of the pituitary are rPAM-3b and rPAM-2. Despite the fact that anterior and neurointermediate pituitary contain a similar distribution of forms of PAM mRNA, the distribution of PAM proteins in the two lobes of the pituitary is quite different. Although integral membrane proteins similar to rPAM-2 and rPAM-3b are major components of anterior pituitary granules, the PAM proteins in the neurointermediate lobe have undergone more extensive endoproteolytic processing, and a 75-kDa protein containing both PHM and PAL domains predominates. The bifunctional PAM precursor undergoes tissue-specific endoproteolytic cleavage reminiscent of the processing of prohormones
Biochemical and spectroscopic properties of DNA-binding zinc fingers: Application of Fmoc-mediated synthesis on PEG-polystyrene
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Human Luteinizing Hormone: Amino-Terminal Sequence Analysis of the β Subunit using [<sup><b>35</b></sup>S]Phenylisothiocyanate
No full textReceptor binding and subunit interaction by the N-terminal (1–15) region of LH/hCG β subunit shown by use of synthetic peptides
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