The hormone-binding site of neurophysins: binding of vasopressin to the N-terminal sub-domain dissected from human MSEL-neurophysin through endopeptidase Lys-C.
Human MSEL-neurophysin has been dissected into two halves by endopeptidase Lys-C, taking advantage of a peculiar Lys59-Ala60 bond. Two sub-domains, N-terminal (1-59) and C-terminal (60-93), have been separated. These sub-domains have been purified by reverse-phase high pressure liquid chromatography and identified by their N-terminal sequences. The N-terminal fragment comprises two chains 1-18 and 19-59, because of the presence of a second lysine residue in position 18, whereas the C-terminal fragment (60-93) is a single chain. Hormone-binding experiments have been carried out using vasopressin or vasopressinyl-Gly-Lys-Arg and testing the ability of the hormone-neurophysin complex to precipitate at pH 3.9 with 10% NaCl. The N-terminal sub-domain precipitates in presence of vasopressin in the same way as native neurophysin whereas the C-terminal sub-domain does not. It can be concluded that the hormone-binding site is located in the 1-59 region of neurophysin.
Michel
G
G
Laboratory of Biological Chemistry, University of Paris VI 96, France.
Chauvet
J
J
Acher
R
R
D000595:Amino Acid Sequence; D001127:Arginine Vasopressin / Q000378:metabolism*; D001665:Binding Sites; D002851:Chromatography, High Pressure Liquid; D010450:Endopeptidases / Q000378:metabolism*; D006801:Humans; D008666:Metalloendopeptidases; D008969:Molecular Sequence Data; D009481:Neurophysins / Q000378:metabolism*; D010121:Oxytocin; D010446:Peptide Fragments / Q000302:isolation & purification / Q000378:metabolism; D011498:Protein Precursors / Q000378:metabolism*; D014667:Vasopressins / Q000378:metabolism*
D009481:Neurophysins; D010446:Peptide Fragments; D011498:Protein Precursors; D014667:Vasopressins; D001127:Arginine Vasopressin; D010121:Oxytocin; D010450:Endopeptidases; D008666:Metalloendopeptidases; C039830:peptidyl-Lys metalloendopeptidase