Stability and functionality of cysteine‐less FOF1 ATP synthase from Escherichia coli
- 17 April 1998
- journal article
- Published by Wiley in FEBS Letters
- Vol. 426 (2) , 217-220
- https://doi.org/10.1016/s0014-5793(98)00337-8
Abstract
All 21 native cysteines in the Escherichia coli FOF1 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 FOF1 is biochemically stable and has functionality similar to wild‐type.Keywords
This publication has 19 references indexed in Scilit:
- Intergenic suppression of the γM23K uncoupling mutation in F0F1 ATP synthase by βGlu-381 substitutions: the role of the β380DELSEED386 segment in energy couplingBiochemical Journal, 1998
- ATP synthase: an electrochemical ransducer with rotatory mechanicsTrends in Biochemical Sciences, 1997
- THE ATP SYNTHASE—A SPLENDID MOLECULAR MACHINEAnnual Review of Biochemistry, 1997
- Energy Coupling, Turnover, and Stability of the F0F1 ATP Synthase Are Dependent on the Energy of Interaction between γ and β SubunitsJournal of Biological Chemistry, 1997
- Mechanisms of Active Transport in the F O F 1 ATP SynthaseThe Journal of Membrane Biology, 1996
- The ATP Synthase γ Subunit.Published by Elsevier ,1995
- Site-Directed Spin Labeling of Membrane ProteinsPublished by Springer Nature ,1994
- A Short Polypeptide Marker Sequence Useful for Recombinant Protein Identification and PurificationBio/Technology, 1988
- Determination of microgram quantities of protein in the presence of milligram levels of lipid with amido black 10B1Analytical Biochemistry, 1985
- Purification and Properties of Reconstitutively Active and Inactive Adenosinetriphosphatase from Escherichia coliProceedings of the National Academy of Sciences, 1974