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Stability and functionality of cysteine‐less F O F 1 ATP synthase from Escherichia coli
Author(s) -
Kuo Phillip H,
Ketchum Christian J,
Nakamoto Robert K
Publication year - 1998
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(98)00337-8
Subject(s) - escherichia coli , atp synthase , cysteine , enzyme , atp hydrolysis , chemistry , biochemistry , chemiosmosis , liposome , hydrolysis , atpase , gene
All 21 native cysteines in the Escherichia coli F O F 1 ATP synthase were replaced by alanines. In isolated E. coli membranes, ATP‐dependent proton pumping, turnover of ATP hydrolysis and steady‐state transition state thermodynamic parameters of the cysteine‐less enzyme were similar to wild‐type. The cysteine‐less enzyme was solubilized in n ‐octyl β‐ d ‐glucopyranoside, purified by affinity chromatography, and reconstituted into pre‐formed liposomes made from E. coli lipids. The properties of the reconstituted, purified enzyme were not significantly different from the membranous enzyme. These data demonstrate that cysteine‐less F O F 1 is biochemically stable and has functionality similar to wild‐type.