Evidence for Multiple Forms of Melatonin Receptor-G-Protein Complexes by Solubilization and Gel Electrophoresis

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25 Citations (Scopus)

Abstract

The daily production of melatonin from the pineal gland influences circadian and seasonal behaviour and physiology. To further understand how melatonin may function, it is important to characterize the receptor and signal transduction systems. Using the detergent digitonin, we were able to solubilize the receptor from the ovine pars tuberalis (PT) membrane. The receptor was isolated as a complex associated with its heterotrimeric G-protein. In the solubilized state, pre-bound I-125-2-iodomelatonin was stable at 4 degrees C, but was displaceable by GTP gamma S. The receptor-G-protein complex could be separated by molecular mass using native polyacrylamide gel electrophoresis. We demonstrate that the receptor-complex has a molecular mass of 525 kDa and differs from solubilized receptor-complexes isolated from either the lizard brain, chicken brain or the ovine hippocampus. Furthermore the receptor complex isolated from the hippocampus had the lowest molecular mass of these tissues (365 kDa) and was found not to be sensitive to GTP gamma S. This may indicate the existence of a distinct non-G-protein coupled form of the receptor.

Original languageEnglish
Pages (from-to)509-515
Number of pages7
JournalJournal of Neuroendocrinology
Volume6
Issue number5
DOIs
Publication statusPublished - Oct 1994

Keywords

  • melatonin
  • pars tuberalis
  • G-protein
  • species
  • electrophoresis
  • ovine pars tuberalis
  • binding-sites
  • hamster-brain
  • lizard-brain
  • localization
  • pituitary
  • cells
  • rat

Cite this

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title = "Evidence for Multiple Forms of Melatonin Receptor-G-Protein Complexes by Solubilization and Gel Electrophoresis",
abstract = "The daily production of melatonin from the pineal gland influences circadian and seasonal behaviour and physiology. To further understand how melatonin may function, it is important to characterize the receptor and signal transduction systems. Using the detergent digitonin, we were able to solubilize the receptor from the ovine pars tuberalis (PT) membrane. The receptor was isolated as a complex associated with its heterotrimeric G-protein. In the solubilized state, pre-bound I-125-2-iodomelatonin was stable at 4 degrees C, but was displaceable by GTP gamma S. The receptor-G-protein complex could be separated by molecular mass using native polyacrylamide gel electrophoresis. We demonstrate that the receptor-complex has a molecular mass of 525 kDa and differs from solubilized receptor-complexes isolated from either the lizard brain, chicken brain or the ovine hippocampus. Furthermore the receptor complex isolated from the hippocampus had the lowest molecular mass of these tissues (365 kDa) and was found not to be sensitive to GTP gamma S. This may indicate the existence of a distinct non-G-protein coupled form of the receptor.",
keywords = "melatonin, pars tuberalis, G-protein, species, electrophoresis, ovine pars tuberalis, binding-sites, hamster-brain, lizard-brain, localization, pituitary, cells, rat",
author = "Perry Barrett and A MACLEAN and Morgan, {Peter John}",
year = "1994",
month = "10",
doi = "10.1111/j.1365-2826.1994.tb00613.x",
language = "English",
volume = "6",
pages = "509--515",
journal = "Journal of Neuroendocrinology",
issn = "0953-8194",
publisher = "Wiley-Blackwell",
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TY - JOUR

T1 - Evidence for Multiple Forms of Melatonin Receptor-G-Protein Complexes by Solubilization and Gel Electrophoresis

AU - Barrett, Perry

AU - MACLEAN, A

AU - Morgan, Peter John

PY - 1994/10

Y1 - 1994/10

N2 - The daily production of melatonin from the pineal gland influences circadian and seasonal behaviour and physiology. To further understand how melatonin may function, it is important to characterize the receptor and signal transduction systems. Using the detergent digitonin, we were able to solubilize the receptor from the ovine pars tuberalis (PT) membrane. The receptor was isolated as a complex associated with its heterotrimeric G-protein. In the solubilized state, pre-bound I-125-2-iodomelatonin was stable at 4 degrees C, but was displaceable by GTP gamma S. The receptor-G-protein complex could be separated by molecular mass using native polyacrylamide gel electrophoresis. We demonstrate that the receptor-complex has a molecular mass of 525 kDa and differs from solubilized receptor-complexes isolated from either the lizard brain, chicken brain or the ovine hippocampus. Furthermore the receptor complex isolated from the hippocampus had the lowest molecular mass of these tissues (365 kDa) and was found not to be sensitive to GTP gamma S. This may indicate the existence of a distinct non-G-protein coupled form of the receptor.

AB - The daily production of melatonin from the pineal gland influences circadian and seasonal behaviour and physiology. To further understand how melatonin may function, it is important to characterize the receptor and signal transduction systems. Using the detergent digitonin, we were able to solubilize the receptor from the ovine pars tuberalis (PT) membrane. The receptor was isolated as a complex associated with its heterotrimeric G-protein. In the solubilized state, pre-bound I-125-2-iodomelatonin was stable at 4 degrees C, but was displaceable by GTP gamma S. The receptor-G-protein complex could be separated by molecular mass using native polyacrylamide gel electrophoresis. We demonstrate that the receptor-complex has a molecular mass of 525 kDa and differs from solubilized receptor-complexes isolated from either the lizard brain, chicken brain or the ovine hippocampus. Furthermore the receptor complex isolated from the hippocampus had the lowest molecular mass of these tissues (365 kDa) and was found not to be sensitive to GTP gamma S. This may indicate the existence of a distinct non-G-protein coupled form of the receptor.

KW - melatonin

KW - pars tuberalis

KW - G-protein

KW - species

KW - electrophoresis

KW - ovine pars tuberalis

KW - binding-sites

KW - hamster-brain

KW - lizard-brain

KW - localization

KW - pituitary

KW - cells

KW - rat

U2 - 10.1111/j.1365-2826.1994.tb00613.x

DO - 10.1111/j.1365-2826.1994.tb00613.x

M3 - Article

VL - 6

SP - 509

EP - 515

JO - Journal of Neuroendocrinology

JF - Journal of Neuroendocrinology

SN - 0953-8194

IS - 5

ER -