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Effects of pH, salt and time on ligand binding properties of overexpressed melanocortin 4 receptor.

Dolby, Viveka LU ; Lundqvist, Andreas; Fröberg, Thomas; Lüllau, Elke; Shaw, Judith; Tjerneld, Folke LU and Cronet, Philippe (2004) In Journal of Biochemical and Biophysical Methods1979-01-01+01:002008-01-01+01:00 58(3). p.195-205
Abstract
The G-protein coupled melanocortin 4 receptor (MC4r) plays an important role in the energy metabolism. We overexpressed the MC4r in CHO cells and performed characterisation studies on the cell membranes to determine functional stability and ligand binding properties of the receptor. The affinity for the ligands [Nle4, Image-Phe7]-αMSH and MTII was lost below pH 6 but could be restored by returning to physiological pH. Increasing NaCl concentration up to 1 M had little influence on the binding of either ligand. At neutral pH, physiological salt concentration and 4 °C the ligand affinity of the receptor was stable for up to 6 days. These findings will facilitate design of purification methods for the receptor.
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Author Keywords: Melanocortin receptor, Ligand binding, G-protein coupled receptor, Antibodies, MTII, NDP-MSH
in
Journal of Biochemical and Biophysical Methods1979-01-01+01:002008-01-01+01:00
volume
58
issue
3
pages
195 - 205
publisher
Elsevier
external identifiers
  • wos:000220766900002
  • pmid:15026206
  • scopus:1542358178
ISSN
0165-022X
DOI
10.1016/j.jbbm.2003.10.007
language
English
LU publication?
yes
id
f4a1e43f-65bb-4e6a-9882-6f1a6935dc37 (old id 121173)
date added to LUP
2007-07-04 16:17:52
date last changed
2017-11-30 16:12:54
@article{f4a1e43f-65bb-4e6a-9882-6f1a6935dc37,
  abstract     = {The G-protein coupled melanocortin 4 receptor (MC4r) plays an important role in the energy metabolism. We overexpressed the MC4r in CHO cells and performed characterisation studies on the cell membranes to determine functional stability and ligand binding properties of the receptor. The affinity for the ligands [Nle4, Image-Phe7]-αMSH and MTII was lost below pH 6 but could be restored by returning to physiological pH. Increasing NaCl concentration up to 1 M had little influence on the binding of either ligand. At neutral pH, physiological salt concentration and 4 °C the ligand affinity of the receptor was stable for up to 6 days. These findings will facilitate design of purification methods for the receptor.},
  author       = {Dolby, Viveka and Lundqvist, Andreas and Fröberg, Thomas and Lüllau, Elke and Shaw, Judith and Tjerneld, Folke and Cronet, Philippe},
  issn         = {0165-022X},
  keyword      = {Author Keywords: Melanocortin receptor,Ligand binding,G-protein coupled receptor,Antibodies,MTII,NDP-MSH},
  language     = {eng},
  number       = {3},
  pages        = {195--205},
  publisher    = {Elsevier},
  series       = {Journal of Biochemical and Biophysical Methods1979-01-01+01:002008-01-01+01:00},
  title        = {Effects of pH, salt and time on ligand binding properties of overexpressed melanocortin 4 receptor.},
  url          = {http://dx.doi.org/10.1016/j.jbbm.2003.10.007},
  volume       = {58},
  year         = {2004},
}