Investigation of the interactions of oxytocin with neurophysins at low pH using carbon-13 nuclear magnetic resonance and carbon-13-labeled hormones

Biochemistry
M BlumensteinV Viswanatha

Abstract

The specifically 13C-labeled (90% 13C-enriched) peptide hormone derivatives [1-hem[2-13C]cystine]oxytocin, [1-hemi[1-13C]cystine]oxytocin, and [2-[-2-13C]tyrosine[-oxytocin and the analogue [3-[2-13C]leucine]oxytocin were prepared by total synthesis and used to study the interactions of the neurohypophyseal hormones with the bovine neurophysins as a function of pH and temperature. Under all conditions, whether high or low pH, the chemical shifts of the labeled carbon atoms of the bound hormones are the same, but they are shifted significantly from their positions in the free hormone. These results indicate that interactions of the side chain and disulfide moieties of the hormone with the neurophysins do not change as a function of pH. At neutral pH and 20--35 degrees C, the labeled atoms of the hormone are in slow exchange (1--5 s-1) with the neurophysins for the above hormone derivatives, but at low pH they are in intermediate or fast exchange depending upon the pH and temperature. At low pH, the dissociation rate constant (koff) is about 100-fold greater than the value at neutral pH, and this increase appears to be due exclusively to the breaking of the salt bridge involving the N-terminal amino group of oxytocin and a side-c...Continue Reading

References

Jun 13, 1959·Nature·J PORATH, P FLODIN

Citations

Dec 17, 2009·The Journal of Organic Chemistry·V J Hruby
Apr 22, 1999·Protein Science : a Publication of the Protein Society·E BreslowR H Haschemeyer
Feb 25, 2003·Bioscience, Biotechnology, and Biochemistry·Takuma UoNobuyoshi Esaki
Nov 1, 1980·International Journal of Peptide and Protein Research·A A Bothner-ByE Breslow

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