caa a22 4500 463208129 CHVBK 20180405153132.0 cr unu---uuuuu 170326e20070201xx s 000 0 eng 10.1007/s10863-006-9066-6 doi (NATIONALLICENCE)springer-10.1007/s10863-006-9066-6 Chemical modification of mono-cysteine mutants allows a more global look at conformations of the ε subunit of the ATP synthase from Escherichia coli [Elektronische Daten] [Sangeeta Ganti, Steven Vik] The ε subunit of the ATP synthase from E. coli undergoes conformational changes while rotating through 360° during catalysis. The conformation of ε was probed in the membrane-bound ATP synthase by reaction of mono-cysteine mutants with 3-N-maleimidyl-propionyl biocytin (MPB) under resting conditions, during ATP hydrolysis, and after inhibition by ADP-AlF3. The relative extents of labeling were quantified after electrophoresis and blotting of the partially purified ε subunit. Residues from the N-terminal β-sandwich domain showed a position-specific pattern of labeling, consistent with prior structural studies. Some residues near the ε-γ interface showed changes up to two-fold if labeling occurred during ATP hydrolysis or after inhibition by ADP-AlF3. In contrast, residues found in the C-terminal α-helices were all labeled to a moderate or high level with a pattern that was consistent with a partially opened helical hairpin. The results indicate that the two C-terminal α-helices do not adopt a fixed conformation under resting conditions, but rather exhibit intrinsic flexibility. Springer Science+Business Media, LLC, 2007 ATP synthase nationallicence ε nationallicence subunit nationallicence Conformational change nationallicence Cysteine nationallicence Mutagenesis nationallicence Chemical modification nationallicence 3-N-maleimidyl-propionyl biocytin (MPB) nationallicence Ganti Sangeeta Department of Biological Sciences, Southern Methodist University, 75275-0376, Dallas, TX, USA aut Vik Steven Department of Biological Sciences, Southern Methodist University, 75275-0376, Dallas, TX, USA aut Journal of Bioenergetics and Biomembranes Kluwer Academic Publishers-Plenum Publishers; http://www.springer-ny.com 39/1(2007-02-01), 99-107 0145-479X 39:1<99 2007 39 10863 https://doi.org/10.1007/s10863-006-9066-6 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s10863-006-9066-6 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Ganti Sangeeta Department of Biological Sciences, Southern Methodist University, 75275-0376, Dallas, TX, USA aut NATIONALLICENCE 700 1- Vik Steven Department of Biological Sciences, Southern Methodist University, 75275-0376, Dallas, TX, USA aut NATIONALLICENCE 773 0- Journal of Bioenergetics and Biomembranes Kluwer Academic Publishers-Plenum Publishers; http://www.springer-ny.com 39/1(2007-02-01), 99-107 0145-479X 39:1<99 2007 39 10863 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer