Solubilization and characterization of calcitonin gene-related peptide binding site from porcine spinal cord
The binding site for calcitonin gene-related peptide (CGRP) was solubilized with 3-[(3-cholamidopropyl)dimethylammonio]-1-propane sulfonate (CHAPS) in an active form from porcine spinal cord. 125I-labeled human alpha-CGRP (125I-CGRP) binding to the solubilized protein was determined by filtration us...
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Published in | Journal of neurochemistry Vol. 50; no. 2; p. 480 |
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Main Authors | , , , , , , , , |
Format | Journal Article |
Language | English |
Published |
England
01.02.1988
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Subjects | |
Online Access | Get more information |
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Summary: | The binding site for calcitonin gene-related peptide (CGRP) was solubilized with 3-[(3-cholamidopropyl)dimethylammonio]-1-propane sulfonate (CHAPS) in an active form from porcine spinal cord. 125I-labeled human alpha-CGRP (125I-CGRP) binding to the solubilized protein was determined by filtration using a GF/B glass filter. The maximal binding activity (approximately 60% of the crude membrane fraction) was obtained with 5 mM CHAPS. 125I-CGRP binding to the solubilized protein was of high affinity, saturability, and high specificity, having KD and Bmax values of 3.69 pM and 338 fmol/mg of protein, respectively. The binding activity was eluted in a single peak with a molecular mass of 400,000 daltons by gel filtration on TSK gel G4000SW. These results suggest that the solubilized protein may be responsible for the specific binding site. |
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ISSN: | 0022-3042 |
DOI: | 10.1111/j.1471-4159.1988.tb02936.x |