Summary
The three-dimensional structure of synthetic human neuropeptide Y in aqueous solution at pH 3.2 and 37°C was determined from two-dimensional 1H NMR data recorded at 600 MHz. A restraint set consisting of 440 interproton distance restraints inferred from NOEs and 11 backbone and 4 side-chain dihedral angle restraints derived from spin-spin coupling constants was used as input for distance geometry calculations in DIANA and simulated annealing and restrained energy minimisation in X-PLOR. The final set of 26 structures is well defined in the region of residues 11–36, with a mean pairwise rmsd of 0.51 Å for the backbone heavy atoms (N, Cα and C) and 1.34 Å for all heavy atoms. Residues 13–36 form an amphipathic α-helix. The N-terminal 10 residues are poorly defined relative to the helical region, although some elements of local structure are apparent. At least one of the three prolines in this N-terminal region co-exists in both cis and trans conformations. An additional set of 24 distances was interpreted as intermolecular distances within a dimer. A combination of distance geometry and restrained simulated annealing yielded a model of the dimer having antiparallel packing of two helical units, whose hydrophobic faces form a well-defined core. Sedimentation equilibrium experiments confirm the observation that neuropeptide Y associates to form dimers and higher aggregates under the conditions of the NMR experiments. Our results therefore support the structural features reported for porcine neuropeptide Y [Cowley, D.J. et al. (1992) Eur. J. Biochem., 205, 1099–1106] rather than the ‘aPP’ fold described previously for human neuropeptide Y [Darbon, H. et al. (1992) Eur. J. Biochem., 209, 765–771].
Similar content being viewed by others
Abbreviations
- NPY:
-
neuropeptide Y
- PP:
-
pancreatic polypeptide
- 1D, 2D:
-
one-, two-dimensional
- NOE:
-
nuclear Overhauser enhancement
- NOESY:
-
2D NOE spectroscopy
- TOCSY:
-
2D total correlation spectroscopy
- E.COSY:
-
exclusive correlation spectroscopy
- HMQC:
-
heteronuclear multiple-quantum coherence
- rmsd:
-
root-mean-square deviation
References
AnilKumar, ErnstR.R. and WüthrichK. (1980) Biochem. Biophys. Res. Commun., 95, 1–6.
BardenJ.A. (1995) Biochem. Biophys. Res. Commun., 215, 264–271.
BaxA., GriffeyR.H. and HawkinsB.L. (1983) J. Am. Chem. Soc., 105, 7188–7190.
BlundellT.L., PittsJ.E., TickleI.J., WoodS.P. and WuC.-W. (1981) Proc. Natl. Acad. Sci. USA, 78, 4175–4179.
BoulangerY., ChenY., CommodariF., SenécalL., LabergeA.-M., FournierA. and St. PierreS. (1995) Int. J. Pept. Protein Res., 45, 86–95.
BraunschweilerL. and ErnstR.R. (1983) J. Magn. Reson. 53, 521–528.
BrooksB.R., BruccoleriR.E., OlafsonB.D., StatesD.J., SwaminathanS. and KarplusM. (1983) J. Comput. Chem., 4, 187–217.
BrüngerA.T. (1992) X-PLOR v. 3.1, A System forX-ray Crystallography and NMR, Yale University, New Haven, CT, U.S.A.
ChangP.J., NoelkenM.E. and KimmelJ.R. (1980) Biochemistry, 19, 1844–1849.
ChazinW.J. and WrightP.E. (1987) Biopolymers, 26, 973–977.
ColmersW.F. and WahlestedtC. (1993) The Biology of Neuropeptide Y and Related Peptides, Humana, Totowa, NJ, U.S.A.
CowleyD.J., HoflackJ.M., PeltonJ.T. and SaudekV. (1992) Eur. J. Biochem., 205, 1099–1106.
DarbonH., BernassauJ.-M., DeleuzeC., ChenuJ., RousselA. and CambillauC. (1992) Eur. J. Biochem., 209, 765–771.
FogolariF., EspositoG., CauciS. and ViglinoP. (1993) J. Magn. Reson., A102, 49–57.
GrayT.S. and MorleyJ.E. (1986) Life Sci., 38, 389–401.
GriesingerC., SørensenO.W. and ErnstR.R. (1987) J. Magn. Reson., 75, 474–492.
Grundemar, L., Sheikh, S.P. and Wahlestedt, C. (1993) In The Biology of Neuropeptide Y and Related Peptides (Eds., Colmers, W.F. and Wahlestedt, C.), Humana, Totowa, NJ, U.S.A., pp. 197–239.
GrundemarL. and HåkansonR. (1994) Trends Pharmacol. Sci., 15, 153–158.
GüntertP., BraunW. and WüthrichK. (1991) J. Mol. Biol., 217, 517–530.
HybertsS.G., MärkiW. and WagnerG. (1987) Eur. J. Biochem., 164, 625–635.
HybertsS.G., GoldbergM.S., HavelT.F. and WagnerG. (1992) Protein Sci., 1, 736–751.
IUPAC-IUB Commission on Biochemical Nomenclature (1970) J. Mol. Biol., 52, 1–17.
LiX., SutcliffeM.J., SchwartzT.W. and DobsonC.M. (1992) Biochemistry, 31, 1245–1253.
MacuraS., HuangY., SuterD. and ErnstR.R. (1981) J. Magn. Reson., 43, 259–281.
MarionD. and WüthrichK. (1983) Biochem. Biophys. Res. Commun., 113, 967–974.
MierkeD.F., DürrH., KesslerH. and JungG. (1992) Eur. J. Biochem., 206, 39–48.
MinakataH., TaylorJ.W., WalkerM.W., MillerR.J. and KaiserE.T. (1989) J. Biol. Chem., 264, 7970–7913.
NilgesM. (1993) Proteins, 17, 297–309.
NoelkenM.E., ChangP.J. and KimmelJ.R. (1980) Biochemistry, 19, 1838–1843.
PallaghyP.K., DugganB.M., PenningtonM.W. and NortonR.S. (1993) J. Mol. Biol., 234, 405–420.
PallaghyP.K., ScanlonM.J., MonksS.A. and NortonR.S. (1995) Biochemistry, 34, 3782–3794.
RuckerS.P. and ShakaA.J. (1989) Mol. Phys., 68, 509–517.
SaudekV. and PeltonJ.T. (1990) Biochemistry, 29, 4509–4515.
SchachmanH.K. (1959) Ultracentrifugation in Biochemistry, Academic Press, New York, NY, U.S.A., pp. 201–247.
WagnerG., BraunW., HavelT.F., SchaumannT., GōN. and WüthrichK. (1987) J. Mol. Biol., 196, 611–639.
WishartD.S., SykesB.D. and RichardsF.M. (1992) Biochemistry, 31, 1647–1651.
WishartD.S. and SykesB.D. (1994) J. Biomol. NMR, 4, 171–180.
WishartD.S., BigamC.G., HolmA., HodgesR.S. and SykesB.D. (1995) J. Biomol. NMR, 5, 67–81.
WüthrichK. (1986) NMR of Proteins and Nucleic Acids, Wiley, New York, NY, U.S.A.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Monks, S.A., Karagianis, G., Howlett, G.J. et al. Solution structure of human neuropeptide Y. J Biomol NMR 8, 379–390 (1996). https://doi.org/10.1007/BF00228141
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00228141