Action of neurohypophysial granule Lys-Arg endopeptidase on synthetic polypeptides comprising the processing sequence of provasopressin-neurophysin.
Neurohypophysial granule Ca(2+)-dependent endopeptidases have been allowed to act on synthetic polypeptides derived from the N-terminal sequence of bovine provasopressin-neurophysin, namely vasopressinyl-glycyl-lysyl-arginyl-alanylamide and vasopressinyl-glycyl-lysyl-arginyl-alanyl-methionyl-serinamide+ ++. Membrane-bound enzymes have been used at pH 5.5 for 16 hr at 37 degrees C. Products have been identified by high-pressure liquid chromatography (HPLC) and by mass spectrometry performed on substances isolated by HPLC. With both substrates, vasopressinyl-Gly-Lys-Arg(OH) has been identified as a product confirming the Lys-Arg specificity previously observed on small peptide fluorogenic substrates. Cleavage yields, however, appear low suggesting that some factors are missing, for example a targeting action of the precursor neurophysin domain to the granule membrane.
Michel
G
G
Laboratory of Biological Chemistry, University of Paris VI.
Rouille
Y
Y
Chauvet
J
J
Acher
R
R
D000595:Amino Acid Sequence; D000818:Animals; D001120:Arginine; D001127:Arginine Vasopressin / Q000378:metabolism*; D002417:Cattle; D003594:Cytoplasmic Granules / Q000201:enzymology; D010450:Endopeptidases / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D066298:In Vitro Techniques; D008239:Lysine; D008969:Molecular Sequence Data; D009481:Neurophysins / Q000378:metabolism*; D010121:Oxytocin; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000378:metabolism; D010904:Pituitary Gland, Posterior / Q000201:enzymology*; D011498:Protein Precursors / Q000378:metabolism*; D051381:Rats; D014667:Vasopressins / Q000378:metabolism*
D009481:Neurophysins; D010446:Peptide Fragments; D011498:Protein Precursors; D014667:Vasopressins; D001127:Arginine Vasopressin; D010121:Oxytocin; D001120:Arginine; D010450:Endopeptidases; D008239:Lysine