Cocaine- and amphetamine-regulated transcript (CART) peptide specific binding in pheochromocytoma cells PC12
Language English Country Netherlands Media print-electronic
Document type Journal Article, Research Support, Non-U.S. Gov't
PubMed
17292884
DOI
10.1016/j.ejphar.2006.12.014
PII: S0014-2999(07)00007-6
Knihovny.cz E-resources
- MeSH
- Models, Biological MeSH
- Cell Differentiation drug effects MeSH
- Cell Membrane metabolism MeSH
- PC12 Cells MeSH
- Time Factors MeSH
- Phenotype MeSH
- Pheochromocytoma MeSH
- Iodine Isotopes metabolism MeSH
- Kinetics MeSH
- Cocaine- and Amphetamine-Regulated Transcript Protein MeSH
- Binding, Competitive MeSH
- Rats MeSH
- Mice, Inbred C57BL MeSH
- Mice MeSH
- Adrenal Gland Neoplasms MeSH
- Nerve Growth Factor pharmacology MeSH
- Neurons metabolism pathology MeSH
- Peptide Fragments metabolism MeSH
- Eating drug effects MeSH
- Nerve Tissue Proteins metabolism pharmacology MeSH
- Iodine Radioisotopes metabolism MeSH
- Protein Binding MeSH
- Dose-Response Relationship, Drug MeSH
- Animals MeSH
- Check Tag
- Rats MeSH
- Male MeSH
- Mice MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- cocaine- and amphetamine-regulated transcript protein (55-102) MeSH Browser
- Iodine Isotopes MeSH
- Cocaine- and Amphetamine-Regulated Transcript Protein MeSH
- Nerve Growth Factor MeSH
- Peptide Fragments MeSH
- Nerve Tissue Proteins MeSH
- Iodine Radioisotopes MeSH
CART (cocaine- and amphetamine-regulated transcript) peptides have been studied for ten years. We report specific binding of 125I-CART(61-102) to the rat adrenal pheochromocytoma PC12 cell line, both intact cells and cell membranes. Saturation binding to intact plated cells resulted in Kd of 0.48+/-0.16 nM and Bmax of 2228+/-529 binding sites/cell. 125I-CART(61-102) was also bound to PC12 cells differentiated using nerve growth factor to the neuronal phenotype with non-specific binding below 20%, and Kd of 1.90+/-0.27 nM and Bmax of 11,194+/-261 binding sites/cell. In competitive binding experiments, CART(61-102), CART(55-102) and di-iodinated CART(61-102) were bound to PC12 cell membranes with Ki in low nM range; their affinity to intact non-differentiated and differentiated cells was in low 10(-8) M range. In order to prove that iodination did not eliminate the pharmacological properties of CART, we tested the biological activity of di-iodinated CART(61-102). It decreased food intake in in vivo feeding experiment on fasted mice in a dose of 1 microg/mouse to the same extent as CART(61-102) in a dose of 0.5 microg/mouse.
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