TY - JOUR
T1 - Structural characterization of the interaction of the δ and α subunits of the Escherichia coli F1F0-ATP synthase by NMR spectroscopy
AU - Wilkens, Stephan
AU - Borchardt, Dan
AU - Weber, Joachim
AU - Senior, Alan E.
PY - 2005/9/6
Y1 - 2005/9/6
N2 - A critical point of interaction between F1 and F0 in the bacterial F1F0-ATP synthase is formed by the α and δ subunits. Previous work has shown that the N-terminal domain (residues 3-105) of the δ subunit forms a 6 α-helix bundle [Wilkens, S., Dunn, S. D., Chandler, J., Dahlquist, F. W., and Capaldi, R. A. (1997) Nat. Struct. Biol. 4, 198-201] and that the majority of the binding energy between δ and F1 is provided by the interaction between the N-terminal 22 residues of the α- and N-terminal domain of the δ subunit [Weber, J., Muharemagic, A., Wilke-Mounts, S., and Senior, A. E. (2003) J. Biol. Chem. 278, 13623-13626]. We have now analyzed a 1:1 complex of the δ-subunit N-terminal domain and a peptide comprising the N-terminal 22 residues of the α subunit by heteronuclear protein NMR spectroscopy. A comparison of the chemical-shift values of α-subunit residues with and without δ N-terminal peptide bound indicates that the binding interface on the N-terminal domain of the δ subunit is formed by α helices 1 and V. NOE cross-peak patterns in 2D l2C/12C-filtered NOESY spectra of the l3C-labeled δ-subunit N-terminal domain in complex with unlabeled peptide verify that residues 8-18 in the α-subunit N-terminal peptide are folded as an α helix when bound to δ N-terminal domain. On the basis of intermolecular contacts observed in 12C/ l3C-filtered NOESY experiments, we describe structural details of the interaction of the δ-subunit N-terminal domain with the α-subunit N-terminal α helix.
AB - A critical point of interaction between F1 and F0 in the bacterial F1F0-ATP synthase is formed by the α and δ subunits. Previous work has shown that the N-terminal domain (residues 3-105) of the δ subunit forms a 6 α-helix bundle [Wilkens, S., Dunn, S. D., Chandler, J., Dahlquist, F. W., and Capaldi, R. A. (1997) Nat. Struct. Biol. 4, 198-201] and that the majority of the binding energy between δ and F1 is provided by the interaction between the N-terminal 22 residues of the α- and N-terminal domain of the δ subunit [Weber, J., Muharemagic, A., Wilke-Mounts, S., and Senior, A. E. (2003) J. Biol. Chem. 278, 13623-13626]. We have now analyzed a 1:1 complex of the δ-subunit N-terminal domain and a peptide comprising the N-terminal 22 residues of the α subunit by heteronuclear protein NMR spectroscopy. A comparison of the chemical-shift values of α-subunit residues with and without δ N-terminal peptide bound indicates that the binding interface on the N-terminal domain of the δ subunit is formed by α helices 1 and V. NOE cross-peak patterns in 2D l2C/12C-filtered NOESY spectra of the l3C-labeled δ-subunit N-terminal domain in complex with unlabeled peptide verify that residues 8-18 in the α-subunit N-terminal peptide are folded as an α helix when bound to δ N-terminal domain. On the basis of intermolecular contacts observed in 12C/ l3C-filtered NOESY experiments, we describe structural details of the interaction of the δ-subunit N-terminal domain with the α-subunit N-terminal α helix.
UR - http://www.scopus.com/inward/record.url?scp=24344456025&partnerID=8YFLogxK
U2 - 10.1021/bi0510678
DO - 10.1021/bi0510678
M3 - Article
C2 - 16128580
AN - SCOPUS:24344456025
VL - 44
SP - 11786
EP - 11794
JO - Biochemistry
JF - Biochemistry
SN - 0006-2960
IS - 35
ER -