Summary
Anti-diabetic sulphonylureas act via high affinity binding sites coupled to K-ATP channels. Endosulfine, an endogenous ligand for these binding sites, was shown to exist in two molecular forms, α and Β, in both the pancreas and the central nervous system. We describe here the isolation, and partial structural characterization of α endosulfine derived from porcine brains by means of a series of chromatography runs and gel electrophoresis. Porcine α endosulfine is a protein with a molecular mass of 13,196 daltons as determined by mass spectrometry and which is N-terminally blocked. Tryptic digestion followed by separation of the fragments by HPLC and automated Edman degradation yielded a total of 72 amino acids in four partial sequences. Comparison of these sequences with that present in the National Biomedical Research Foundation protein data bank indicated a 82% identity with a 112-amino acid protein with a molecular mass of 12,353 daltons called ‘cyclic AMP-regulated phosphoprotein-19’, isolated from the bovine brain as a substrate for protein kinase A.
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Abbreviations
- BSA:
-
Bovine serum albumin
- PAGE:
-
polyacrylamide gel electrophoresis
- ARPP-19:
-
cyclic AMP-regulated phosphoprotein-19
- TFA:
-
trifluoroacetic acid
- PTH:
-
phenylthiohydantoin
- CNS:
-
central nervous system
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Virsolvy-Vergine, A., Salazar, G., Sillard, R. et al. Endosulfine, endogenous ligand for the sulphonylurea receptor: isolation from porcine brain and partial structural determination of the α form. Diabetologia 39, 135–141 (1996). https://doi.org/10.1007/BF00403955
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DOI: https://doi.org/10.1007/BF00403955