Detection of binding activity and biologic effect of a novel alpha-melanocyte-stimulating hormone analogue.
- Author:
Ying YING
1
;
Xiao-Peng LAN
;
Ye-Ping TIAN
Author Information
1. Department of Laboratory Examination, Fuzhou General Hospital, Nanjing Military Command, Fuzhou 350025, China.
- Publication Type:Journal Article
- MeSH:
Amino Acid Sequence;
Binding, Competitive;
Cell Line;
Cell Line, Tumor;
Cyclic AMP;
metabolism;
Genetic Vectors;
Humans;
Iodine Radioisotopes;
Kinetics;
Molecular Sequence Data;
Plasmids;
genetics;
Radioligand Assay;
Receptor, Melanocortin, Type 1;
agonists;
genetics;
metabolism;
Receptors, Corticotropin;
agonists;
genetics;
metabolism;
Receptors, Melanocortin;
agonists;
genetics;
metabolism;
Transfection;
Tritium;
alpha-MSH;
analogs & derivatives;
chemistry;
metabolism;
pharmacology
- From:
Acta Pharmaceutica Sinica
2007;42(3):269-273
- CountryChina
- Language:Chinese
-
Abstract:
Binding activity and biologic effect of a novel alpha-melanocyte-stimulating hormone analogue were tested on cells transiently expressing the human melanocortin-1 (MC1), MC3, MC4, and MC5 receptors. The human MC1 and MC5 receptor genes were cloned into the expression vector pcDNA3. 1/ myc-his(-) B. The vectors were transferred to HEK-293 cells by the calcium phosphate method. Stable receptor populations were generated using G418 selection (900 microg x mL(-1)) for subsequent bioassay analysis. K(i) values of the novel alpha-MSH analogue for MC1, MC3, MC4, and MC5 receptors were obtained in competition with [125I]-NDP-MSH for binding studies. The cyclic AMP level was tested by using [3H]-cyclic AMP kit. It is showed that K(i) values of the novel alpha-MSH analogue for MC1, MC3, MC4, and MC5 receptors were (0.159 +/- 0.040), (35.430 +/- 6.743), (19.293 +/- 2.780) and (2.230 +/- 0.670) nmol L(-1), respectively. Its EC50 values for MC1, MC3, MC4, and MC5 receptors were (0.45 +/- 0.07), (7.80 +/- 0.65), (2.55 +/- 0.23) and (0.33 +/- 0.09) nmol L(-1), respectively. In these tests, the novel alpha-MSH analogue is a MC1R and MC5R selective agonist.
