title: Solution structure of an analogue of vasoactive intestinal peptide as determined by two-dimensional NMR and circular dichroism spectroscopies and constrained molecular dynamics.
abstract: Structures have been determined for a potent analogue of vasoactive intestinal peptide (VIP), Ac-[Lys12, Lys14, Nle17, Val26, Thr28]VIP (VIP'), in methanol/water solutions. In CD studies, both VIP and VIP' were helical in methanol/water, with the percentage of alpha-helix increasing with percentage methanol. The pH had little effect on the structure. Complete 1H NMR assignments were made for VIP' in 25% methanol at pH 4 and 6 and in 50% methanol at pH 6, using two-dimensional COSY, NOESY, and relay-COSY experiments. There were no widespread changes in chemical shifts between the samples at pH 4 and 6; however, widespread changes were observed between the samples in 25% and 50% methanol. Complete sets of NOEs were obtained for VIP' in 25% methanol, pH 4, and in 50% methanol, pH 6. These NOEs were converted into distance constraints and applied in molecular dynamics and energy minimization calculations using the program CHARMM. A set of low-energy structures was obtained for VIP' in each solvent system. In 25% methanol, VIP' has two helical segments at residues 9-17 and 23-28. The remainder of the structure is not well determined. In 50% methanol, residues 8-26 form a regular, well-defined alpha-helix and residues 5-8 form a type III beta-turn. The remaining residues are not ordered. These structural assessments agree with the CD data. In the lowest energy structure in 50% methanol, the side chains of Asp3, Phe6, Thr7, and Tyr10 are clustered together--these residues are conserved throughout the family of peptide hormones homologous to VIP.
authors.affiliation: Department of Physical Chemistry, Hoffmann-La Roche Inc., Nutley, New Jersey 07110.
authors.forename: D C
authors.initials: DC
authors.lastname: Fry
authors.forename: V S
authors.initials: VS
authors.lastname: Madison
authors.forename: D R
authors.initials: DR
authors.lastname: Bolin
authors.forename: D N
authors.initials: DN
authors.lastname: Greeley
authors.forename: V
authors.initials: V
authors.lastname: Toome
authors.forename: B B
authors.initials: BB
authors.lastname: Wegrzynski
mesh_terms: D002942:Circular Dichroism; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D008958:Models, Molecular; D011487:Protein Conformation; D013816:Thermodynamics; D014660:Vasoactive Intestinal Peptide / Q000031:analogs & derivatives*
chemical_list: C060055:vasoactive intestinal peptide, Ac-(Lys(12,14)-Nle(17)-Val(26)-Thr(28))-; D014660:Vasoactive Intestinal Peptide