Effects of pH, salt and time on ligand binding properties of overexpressed melanocortin 4 receptor.
(2004) In Journal of Biochemical and Biophysical Methods 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.
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/121173
- author
- Dolby, Viveka LU ; Lundqvist, Andreas ; Fröberg, Thomas ; Lüllau, Elke ; Shaw, Judith ; Tjerneld, Folke LU and Cronet, Philippe
- organization
- publishing date
- 2004
- 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 Methods
- 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
- 2016-04-01 15:38:15
- date last changed
- 2022-01-28 06:21:58
@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}}, keywords = {{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 Methods}}, 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}}, doi = {{10.1016/j.jbbm.2003.10.007}}, volume = {{58}}, year = {{2004}}, }