1H and 15N NMR characterization of free and bound states of an amphiphilic peptide interacting with calmodulin
| Autor: | Prêcheur, B., Munier, Hélène, Mispelter, Joël, Bârzu, O., Craescu, Constantin |
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| Přispěvatelé: | Génétique moléculaire et hématologie, Institut National de la Santé et de la Recherche Médicale (INSERM), Biochimie des Régulations Cellulaires, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Biophysique moléculaire, Institut Curie [Paris], Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS) |
| Jazyk: | angličtina |
| Rok vydání: | 1992 |
| Předmět: |
Models
Molecular Magnetic Resonance Spectroscopy Nitrogen Isotopes [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry Molecular Biology/Structural Biology [q-bio.BM] Protein Conformation Swine Circular Dichroism Molecular Sequence Data Calmodulin Solubility Animals Amino Acid Sequence Peptides Hydrogen Protein Binding |
| Zdroj: | Biochemistry Biochemistry, 1992, 31 (1), pp.229-36 Biochemistry, American Chemical Society, 1992, 31 (1), pp.229-36 |
| ISSN: | 0006-2960 1520-4995 |
| Popis: | International audience; A peptide of 17 amino acid residues Ac-L-K-W-K-K-L-L-K-L-L-K-K-L-L-K-L-G-NH2, designed to form an amphiphilic basic alpha-helix [DeGrado, W.F., Prendergast, F. G., Wolfe, H. R., Jr., & Cox, J. A. (1985) J. Cell. Biochem. 29, 83-93], was labeled with 15N at positions 1, 7, 9, and 10. Homo- and heteronuclear NMR techniques were used to characterize the conformational changes of the peptide when it binds to calmodulin in the presence of Ca2+ ions. The spectrum of the free peptide in aqueous solution at pH 6.3 and 298 K was completely assigned by a combined application of several two-dimensional proton NMR methods. Analysis of the short- and medium-range NOE connectivities and of the secondary chemical shifts indicated that the peptide populates, to a significant extent, an alpha-helix conformational state, in agreement with circular dichroism measurements under similar physicochemical conditions. 15N-edited 1D spectra and 15N(omega 2)-half-filtered two-dimensional NMR experiments on the peptide in a 1:1 complex with calmodulin allowed assignment of half of the amide proton resonances and three C alpha H resonances of the bound peptide. The observed NOE connectivities between the peptide backbone protons are indicative of a stable helical secondary structure spanning at least the fragment L1-K11. The equilibrium and dynamic NMR parameters of the bound peptide are discussed in terms of a molecular interaction model. |
| Databáze: | OpenAIRE |
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