Amino acid sequence and properties of vasopressin-associated elephant neurophysin.
The primary structure of an elephant neurophysin, homologous to vasopressin-associated neurophysins, is reported. The protein contains a Tyr for Asn substitution at position 75, a position in direct contact with residues 77 and 78 of the monomer-monomer interface. This Tyr residue therefore serves as a potential reporter of the path involved in the long-range linkage between peptide binding and dimerization in this system. NMR studies of the protein in unliganded and liganded states demonstrated normal dimerization properties and the expected increase in dimerization associated with binding peptide. In keeping with an elevated pKa of 11.1 assigned to Tyr-75 by UV spectrophotometric titration, the NMR signals from the 3,5 and 2,6 ring protons of Tyr-75 were shifted 0.3 and 0.2 ppm upfield, respectively, relative to their positions in small peptides, indicating significant shielding and/or hydrogen bonding. The Tyr-75 ring proton signals narrowed slightly, with no discernible change in chemical shift, on conversion from dimer to monomer in the unliganded state. Ring protons of Tyr-49, distant from the monomer-monomer interface, but adjacent to the peptide-binding site, were markedly perturbed by dimerization, in accord with their behavior in bovine neurophysins. The results suggest that the secondary and tertiary structure of the region 75-78 is largely unchanged by dimerization, and argue against an important role for this region in dimerization-mediated conformational changes that alter the binding site in the unliganded state.(ABSTRACT TRUNCATED AT 250 WORDS)
Huang
H B
HB
Department of Biochemistry, Cornell University Medical College, New York, NY.
Wellner
D
D
Naudé
R
R
Oelofsen
W
W
Oosthuizen
M M
MM
Breslow
E
E
D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D002848:Chromatography, DEAE-Cellulose; D002851:Chromatography, High Pressure Liquid; D002942:Circular Dichroism; D004606:Elephants; D006863:Hydrogen-Ion Concentration; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D009481:Neurophysins / Q000737:chemistry* / Q000302:isolation & purification; D010904:Pituitary Gland, Posterior / Q000737:chemistry*; D011487:Protein Conformation; D017386:Sequence Homology, Amino Acid; D013056:Spectrophotometry, Ultraviolet; D014443:Tyrosine; D014667:Vasopressins / Q000737:chemistry*
D009481:Neurophysins; D014667:Vasopressins; D014443:Tyrosine