TY - JOUR
T1 - The ε subunit of Escherichia coli coupling factor 1 is required for its binding to the cytoplasmic membrane
AU - Sternweis, P. C.
N1 - Copyright:
Copyright 2017 Elsevier B.V., All rights reserved.
PY - 1978
Y1 - 1978
N2 - The coupling factor, F1-ATPase of Escherichia coli (ECF1) contains five different subunits, α, β, γ, δ and ε. Properties of δ-deficient ECF1 have previously been described. F1ATPase containing only the α, β, and γ subunits was prepared from E.coli by passage of δ-deficient ECF1 through an affinity column containing immobilized antibodies to the ε subunit. The δ,ε-deficient enzyme has normal ATPase activity but cannot bind to ECF1-depleted membrane vesicles. Both the δ and ε subunits are required for the binding of δ,ε-deficient ECF1, to membranes and the restoration of oxidative phosphorylation. Either δ or ε will bind to the deficient enzyme to form a four-subunit complex. Neither four-subunit enzyme binds to depleted membranes. The ε subunit, does, however, slightly improve the binding affinity between δ and δ-deficient enzyme suggesting a possible interaction between the two subunits. Neither subunit binds to trypsin-treated ECF1, which contains only the α and β subunits. A role for γ in the binding of ε to F1 is suggested. ε does not bind to ECF1-depleted membranes. Therefore, the in vitro reconstitution of depleted membranes requires an initial complex formation between ε and the rest of ECF1 prior to membrane attachment. Reconstitution experiments indicate that only one ε is required per functional ECF1 molecule.
AB - The coupling factor, F1-ATPase of Escherichia coli (ECF1) contains five different subunits, α, β, γ, δ and ε. Properties of δ-deficient ECF1 have previously been described. F1ATPase containing only the α, β, and γ subunits was prepared from E.coli by passage of δ-deficient ECF1 through an affinity column containing immobilized antibodies to the ε subunit. The δ,ε-deficient enzyme has normal ATPase activity but cannot bind to ECF1-depleted membrane vesicles. Both the δ and ε subunits are required for the binding of δ,ε-deficient ECF1, to membranes and the restoration of oxidative phosphorylation. Either δ or ε will bind to the deficient enzyme to form a four-subunit complex. Neither four-subunit enzyme binds to depleted membranes. The ε subunit, does, however, slightly improve the binding affinity between δ and δ-deficient enzyme suggesting a possible interaction between the two subunits. Neither subunit binds to trypsin-treated ECF1, which contains only the α and β subunits. A role for γ in the binding of ε to F1 is suggested. ε does not bind to ECF1-depleted membranes. Therefore, the in vitro reconstitution of depleted membranes requires an initial complex formation between ε and the rest of ECF1 prior to membrane attachment. Reconstitution experiments indicate that only one ε is required per functional ECF1 molecule.
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M3 - Article
C2 - 147871
AN - SCOPUS:0018200296
SN - 0021-9258
VL - 253
SP - 3123
EP - 3128
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
IS - 9
ER -