Membrane-induced structure of the mammalian tachykinin neuropeptide gamma.
Neuropeptide gamma (NPgamma) is a neurokinin-2 (NK-2) receptor selective agonist, which plays an important role in mediation of asthma and elicits a wide range of biological responses like bronchoconstriction, vasodepression and regulation of endocrine functions. The structure determination of this peptide agonist is important in understanding the molecular basis of peptide ligand recognition by the receptor and for rational drug design. In the present study we report the solution structure of NPgamma characterized by circular dichroism (CD) spectropolarimetry and 2D (1)H NMR spectroscopy in both aqueous and membrane mimetic solvents. Effect of calcium ions on the conformation of NPgamma was also studied using CD spectropolarimetry. Sequence-specific resonance assignments of protons have been made with the aid of correlation spectroscopy experiments and nuclear Overhauser effect spectroscopy experiments. The distance constraints obtained from the NMR data have been utilized to generate a family of structures, which have been refined using restrained energy minimization and dynamics. These data show that in water NPgamma prefers to be in an extended chain conformation whereas a helical conformation is induced in the central core and the C-terminal region of the peptide (K13-M21) in the presence of perdeuterated dodecylphosphocholine micelles, a membrane model system. A type II' beta turn from H9 to R11 precedes the helical core in the C-terminus of NPgamma. N-terminus of NPgamma also displays some degree of order and a possible turn structure. Conformation adopted by NPgamma in presence of lipid micelles represents a structural motif typical of NK-2 selective agonists and is similar to that observed for Neurokinin A in hydrophobic environment. The observed conformational features have been correlated to the binding ability and biological activity of NPgamma.
Chandrashekar
Indu R
IR
School of Life Sciences, Jawaharlal Nehru University, New Delhi 110067, India.
Dike
Anjali
A
Cowsik
Sudha M
SM
D020816:Amino Acid Motifs; D000818:Animals; D002118:Calcium / Q000737:chemistry; D002462:Cell Membrane / Q000737:chemistry; D002942:Circular Dichroism; D006801:Humans; D007477:Ions; D008024:Ligands; D008055:Lipids / Q000737:chemistry; D009682:Magnetic Resonance Spectroscopy; D008823:Micelles; D008958:Models, Molecular; D010446:Peptide Fragments / Q000737:chemistry*; D010455:Peptides / Q000737:chemistry; D011487:Protein Conformation; D017510:Protein Folding; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D011522:Protons; D012984:Software; D013053:Spectrophotometry; D015320:Tachykinins / Q000737:chemistry*
D007477:Ions; D008024:Ligands; D008055:Lipids; D008823:Micelles; D010446:Peptide Fragments; D010455:Peptides; D011522:Protons; D015320:Tachykinins; C059905:tachykinin neuropeptide gamma; D002118:Calcium