TY - JOUR
T1 - F1F0-ATP synthase
T2 - Binding of δ subunit to a 22-residue peptide mimicking the N-terminal region of α subunit
AU - Weber, Joachim
AU - Muharemagic, Alma
AU - Wilke-Mounts, Susan
AU - Senior, Alan E.
PY - 2003/4/18
Y1 - 2003/4/18
N2 - The stator in F1F0-ATP synthase resists strain generated by rotor torque. In Escherichia coli the b2δ subunit complex comprises the stator, bound to subunit a in F0 and to α3β3 hexagon of F1. Proteolysis and cross-linking had suggested that N-terminal residues of a subunit are involved in binding δ. Here we demonstrate that a synthetic peptide consisting of the first 22 residues of α ("αN1-22") binds specifically to isolated wild-type δ subunit with high affinity (Kd = 130 nM), accounting for a major portion of the binding energy when δ-depleted F1 and isolated δ bind together (Kd = 1.4 nM). Stoichiometry of binding of αN1-22 to δ at saturation was 1/1, showing that in intact F1F0-ATP synthase only one of the three α subunits is involved in δ binding. When αN1-22 was incubated with δ subunits containing mutations in helices 1 or 5 on the F1-binding face of δ, peptide binding was impaired as was binding of δ-depleted F1. Residues α6-18 are predicted to be helical, and a potential helix capping box occurs at residues α3-8. Circular dichroism measurements showed that αN1-22 had significant helical content. Hypothetically a helical region of residues αN1-22 packs with helices 1 and 5 on the F1-binding face of δ, forming the α/δ interface.
AB - The stator in F1F0-ATP synthase resists strain generated by rotor torque. In Escherichia coli the b2δ subunit complex comprises the stator, bound to subunit a in F0 and to α3β3 hexagon of F1. Proteolysis and cross-linking had suggested that N-terminal residues of a subunit are involved in binding δ. Here we demonstrate that a synthetic peptide consisting of the first 22 residues of α ("αN1-22") binds specifically to isolated wild-type δ subunit with high affinity (Kd = 130 nM), accounting for a major portion of the binding energy when δ-depleted F1 and isolated δ bind together (Kd = 1.4 nM). Stoichiometry of binding of αN1-22 to δ at saturation was 1/1, showing that in intact F1F0-ATP synthase only one of the three α subunits is involved in δ binding. When αN1-22 was incubated with δ subunits containing mutations in helices 1 or 5 on the F1-binding face of δ, peptide binding was impaired as was binding of δ-depleted F1. Residues α6-18 are predicted to be helical, and a potential helix capping box occurs at residues α3-8. Circular dichroism measurements showed that αN1-22 had significant helical content. Hypothetically a helical region of residues αN1-22 packs with helices 1 and 5 on the F1-binding face of δ, forming the α/δ interface.
UR - http://www.scopus.com/inward/record.url?scp=0038190861&partnerID=8YFLogxK
U2 - 10.1074/jbc.C300061200
DO - 10.1074/jbc.C300061200
M3 - Article
C2 - 12595534
AN - SCOPUS:0038190861
SN - 0021-9258
VL - 278
SP - 13623
EP - 13626
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
IS - 16
ER -