Interaction of bovine neurophysin with oxytocin and vasopressin measured by temperature-jump relaxation.
The interaction between bovine neurophysins I and II and oxytocin and vasopressin was measured using temperature-jump relaxation. A single relaxation time in the 10 to 90 ms range was noted for each solution. This time depended upon the concentration of both neurophysin and hormone and increased with increasing pH. The formation rate constants (+/- SE) for the interaction of neurophysin I dimer with the protonated form of oxytocin and vasopressin at pH 7.4 in 0.1 M KNO3 and 25 degrees C were 2.8 (+/- 0.4) x 10(6) and 2.3 (+/- 0.5) x 10(6) M-1 s-1, respectively; the dissociation rate constants were 11 and 15 s-1, respectively.For neurophysin II dimer, formation rate constants were 6.0 (+/- 0.2) x 10(6) and 2.4 (+/- 0.3) x 10(6) M-1 s-1; dissociation rate constants were 24 and 16 s-1 for oxytocin and vasopressin, respectively. Formation rate constants for the interaction of neurophysin monomer with protinated hormone were approximatley an order of magniment with circular dichorism and pH titration studies which indicate that this interaction occurs between a negatively charged carboxyl group on the neurophysin and a protonated alpha-amino group on the hormone. Our results indicate that oxytocin and vasopressin bind with greater affinity to the neurophysin dimer than the monomer and that the binding of oxytocin and vasopressin in neurophysin dimer than the monomer and that the binding of oxytocin and vasopressin in neurophysin dimer at pH 7.4 and concentrations between 10(-4) and 10(-5) M is nearly identical for each hormone.
D000818:Animals; D001665:Binding Sites; D002417:Cattle; D004591:Electrophoresis, Polyacrylamide Gel; D007700:Kinetics; D008236:Lypressin; D046911:Macromolecular Substances; D008433:Mathematics; D009481:Neurophysins / Q000302:isolation & purification; D010121:Oxytocin; D011485:Protein Binding; D013696:Temperature; D014667:Vasopressins / Q000031:analogs & derivatives
D046911:Macromolecular Substances; D009481:Neurophysins; D014667:Vasopressins; D010121:Oxytocin; D008236:Lypressin