title: NMR solution structure of a complex of calmodulin with a binding peptide of the Ca2+ pump.
abstract: The three-dimensional structure of the complex between calmodulin (CaM) and a peptide corresponding to the N-terminal portion of the CaM-binding domain of the plasma membrane calcium pump, the peptide C20W, has been solved by heteronuclear three-dimensional nuclear magnetic resonance (NMR) spectroscopy. The structure calculation is based on a total of 1808 intramolecular NOEs and 49 intermolecular NOEs between the peptide C20W and calmodulin from heteronuclear-filtered NOESY spectra and a half-filtered experiment, respectively. Chemical shift differences between free Ca(2+)-saturated CaM and its complex with C20W as well as the structure calculation reveal that C20W binds solely to the C-terminal half of CaM. In addition, comparison of the methyl resonances of the nine assigned methionine residues of free Ca(2+)-saturated CaM with those of the CaM/C20W complex revealed a significant difference between the N-terminal and the C-terminal domain; i.e., resonances in the N-terminal domain of the complex were much more similar to those reported for free CaM in contrast to those in the C-terminal half which were significantly different not only from the resonances of free CaM but also from those reported for the CaM/M13 complex. As a consequence, the global structure of the CaM/C20W complex is unusual, i.e., different from other peptide calmodulin complexes, since we find no indication for a collapsed structure. The fine modulation in the peptide protein interface shows a number of differences to the CaM/M13 complex studied by Ikura et al. [Ikura, M., Clore, G. M., Gronenborn, A. M., Zhu, G., Klee, C. B., and Bax, A. (1992) Science 256, 632-638]. The unusual binding mode to only the C-terminal half of CaM is in agreement with the biochemical observation that the calcium pump can be activated by the C-terminal half of CaM alone [Guerini, D., Krebs, J., and Carafoli, E. (1984) J. Biol. Chem. 259, 15172-15177].
authors.affiliation: Institute of Organic Chemistry, University of Frankfurt, Germany.
authors.forename: B
authors.initials: B
authors.lastname: Elshorst
authors.forename: M
authors.initials: M
authors.lastname: Hennig
authors.forename: H
authors.initials: H
authors.lastname: Försterling
authors.forename: A
authors.initials: A
authors.lastname: Diener
authors.forename: M
authors.initials: M
authors.lastname: Maurer
authors.forename: P
authors.initials: P
authors.lastname: Schulte
authors.forename: H
authors.initials: H
authors.lastname: Schwalbe
authors.forename: C
authors.initials: C
authors.lastname: Griesinger
authors.forename: J
authors.initials: J
authors.lastname: Krebs
authors.forename: H
authors.initials: H
authors.lastname: Schmid
authors.forename: T
authors.initials: T
authors.lastname: Vorherr
authors.forename: E
authors.initials: E
authors.lastname: Carafoli
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000252:Calcium-Transporting ATPases / Q000737:chemistry* / Q000378:metabolism*; D002147:Calmodulin / Q000737:chemistry* / Q000378:metabolism*; D002417:Cattle; D018360:Crystallography, X-Ray; D046911:Macromolecular Substances; D008958:Models, Molecular; D008969:Molecular Sequence Data; D019906:Nuclear Magnetic Resonance, Biomolecular; D010446:Peptide Fragments / Q000737:chemistry* / Q000378:metabolism*; D011485:Protein Binding; D017510:Protein Folding; D017433:Protein Structure, Secondary; D012996:Solutions; D014982:Xenopus laevis
chemical_list: D002147:Calmodulin; D046911:Macromolecular Substances; D010446:Peptide Fragments; D012996:Solutions; D000252:Calcium-Transporting ATPases