Characterization of high-affinity melatonin binding sites in purified cell nuclei of rat liver - PubMed (original) (raw)
Characterization of high-affinity melatonin binding sites in purified cell nuclei of rat liver
D Acuña-Castroviejo et al. J Pineal Res. 1994 Mar.
Abstract
High-affinity 2-125I-iodomelatonin binding sites in homogenates of purified cell nuclei from rat liver were localized and characterized using biochemical binding techniques. Binding at these sites was found to be rapid, reversible, saturable, and to demonstrate pharmacological selectivity. At 0 degrees C, binding reached equilibrium in about 10 min. Scatchard analysis of the data at equilibrium revealed a single class of binding sites with a dissociation constant of KD = 190 +/- 47 pM, Bmax = 9.8 +/- 0.6 fmol/mg protein, and a Hill coefficient of nH = 1.02 +/- 0.034. Kinetic analysis of the association and dissociation curves indicated a kinetic KD = 148 +/- 41 pM, which is in good agreement with the value obtained at equilibrium. The specific binding of 2-125I-iodomelatonin (45 pM) (0.51 +/- 0.04 fmol/mg protein) was significantly improved (0.79 +/- 0.04 fmol/mg protein) when the homogenates of purified liver cell nuclei were preincubated with DNase (2 micrograms/ml at 37 degrees C for 20 min) before being used in binding experiments. After the addition of either proteinase K or trichloroacetic acid to DNase-treated purified cell nuclear homogenates, the specific binding disappeared. This suggests that the specific binding of 2-125I-iodomelatonin in liver cell nuclei is associated with nuclear protein. Competition experiments show that N-acetyl-serotonin (Ki = 81.3 nM) was more potent than 5-hydroxytryptamine (Ki > 1 microM) and 5-methoxytryptamine (Ki >> 10 microM) in inhibiting 2-125I-iodomelatonin binding (Ki melatonin = 146 pM). These data indicate that specific 2-125I-iodomelatonin binding sites exist in the cell nuclei of rat liver, and that they may comprise a locus for the intracellular action of melatonin. The correlation between the KD and Bmax values with melatonin concentrations in nuclei suggest that these binding sites may be a physiological melatonin receptor, which could explain the described genomic effects of the pineal hormone.
Similar articles
- Melatonin receptors in purified cell nuclei of liver.
Acuña-Castroviejo D, Pablos MI, Menendez-Pelaez A, Reiter RJ. Acuña-Castroviejo D, et al. Res Commun Chem Pathol Pharmacol. 1993 Nov;82(2):253-6. Res Commun Chem Pathol Pharmacol. 1993. PMID: 8303096 - Characterization and regulation of the human ML1A melatonin receptor stably expressed in Chinese hamster ovary cells.
Witt-Enderby PA, Dubocovich ML. Witt-Enderby PA, et al. Mol Pharmacol. 1996 Jul;50(1):166-74. Mol Pharmacol. 1996. PMID: 8700109 - Characterization of the chicken brain melatonin-binding protein using iodinated and tritiated ligands.
Kennaway DJ, Hugel HM, Rowe SA. Kennaway DJ, et al. J Pineal Res. 1994 Oct;17(3):137-48. doi: 10.1111/j.1600-079x.1994.tb00125.x. J Pineal Res. 1994. PMID: 7897586 - Binding sites for 2-[125I]iodomelatonin in the adrenal gland.
Brown GM, Pang CS, Pang SF. Brown GM, et al. Biol Signals. 1994 Mar-Apr;3(2):91-8. doi: 10.1159/000109530. Biol Signals. 1994. PMID: 7951652 Review. - 2-[125I]iodomelatonin binding sites in the lung and heart: a link between the photoperiodic signal, melatonin, and the cardiopulmonary system.
Pang CS, Brown GM, Tang PL, Cheng KM, Pang SF. Pang CS, et al. Biol Signals. 1993 Jul-Aug;2(4):228-36. doi: 10.1159/000109496. Biol Signals. 1993. PMID: 8205166 Review.
Cited by
- PORTAL: pilot study on the safety and tolerance of preoperative melatonin application in patients undergoing major liver resection: a double-blind randomized placebo-controlled trial.
Schemmer P, Nickkholgh A, Schneider H, Sobirey M, Weigand M, Koch M, Weitz J, Büchler MW. Schemmer P, et al. BMC Surg. 2008 Jan 23;8:2. doi: 10.1186/1471-2482-8-2. BMC Surg. 2008. PMID: 18215253 Free PMC article. Clinical Trial. - Melatonin and nitric oxide: two required antagonists for mitochondrial homeostasis.
Acuña-Castroviejo D, Escames G, López LC, Hitos AB, León J. Acuña-Castroviejo D, et al. Endocrine. 2005 Jul;27(2):159-68. doi: 10.1385/ENDO:27:2:159. Endocrine. 2005. PMID: 16217129 Review. - Eliminating animal facility light-at-night contamination and its effect on circadian regulation of rodent physiology, tumor growth, and metabolism: a challenge in the relocation of a cancer research laboratory.
Dauchy RT, Dupepe LM, Ooms TG, Dauchy EM, Hill CR, Mao L, Belancio VP, Slakey LM, Hill SM, Blask DE. Dauchy RT, et al. J Am Assoc Lab Anim Sci. 2011 May;50(3):326-36. J Am Assoc Lab Anim Sci. 2011. PMID: 21640027 Free PMC article. - Phytomelatonin: assisting plants to survive and thrive.
Reiter RJ, Tan DX, Zhou Z, Cruz MH, Fuentes-Broto L, Galano A. Reiter RJ, et al. Molecules. 2015 Apr 22;20(4):7396-437. doi: 10.3390/molecules20047396. Molecules. 2015. PMID: 25911967 Free PMC article. Review. - From Chronodisruption to Sarcopenia: The Therapeutic Potential of Melatonin.
Fernández-Martínez J, Ramírez-Casas Y, Yang Y, Aranda-Martínez P, Martínez-Ruiz L, Escames G, Acuña-Castroviejo D. Fernández-Martínez J, et al. Biomolecules. 2023 Dec 12;13(12):1779. doi: 10.3390/biom13121779. Biomolecules. 2023. PMID: 38136651 Free PMC article. Review.
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources