In vivo affinity label of a protein expressed in Escherichia coli
Open Access
- 27 December 1993
- journal article
- Published by Wiley in FEBS Letters
- Vol. 336 (2) , 231-235
- https://doi.org/10.1016/0014-5793(93)80809-9
Abstract
When Tyr‐307 of the β subunit of f1 ‐ATPase from a thennophilic Bacillus strain PS3 is replaced by cysteine and expressed in Escherichia coli cells, about a half population of the mutant β subunit are labeled by Coenzyme A at Cys‐307 through a disulfide bond which is cleavable by reducing treatment. The mutant β subunit can be reconstituted into the α3β3, complex of which ATPase activity is stimulated two‐fold by reducing treatment either prior or after reconstitution. Since Tyr‐307 has been supposed to be located at one of subdomains which form the ATP binding site of the β subunit, Coenzyme A binds to the mutant β subunit as an AT(D)P analogue in E. coli cells and then covalently attaches to Cys‐307.Keywords
This publication has 26 references indexed in Scilit:
- Structure and Mechanism of F 0 F 1 ‐Type ATP Synthases and ATPasesPublished by Wiley ,1991
- The Proton-Translocating ATPase of Escherichia ColiAnnual Review of Biophysics, 1990
- Aromatic rings of tyrosine residues at adenine nucleotide binding sites of the β subunits of F1-ATPase are not necessary for ATPase activityBiochemical and Biophysical Research Communications, 1990
- ATP SYNTHASE (H+-ATPase): Results by Combined Biochemical and Molecular Biological ApproachesAnnual Review of Biochemistry, 1989
- α3β3 complex of thermophilic ATP synthase Catalysis without the γ‐subunitFEBS Letters, 1989
- Sequence and over-expression of subunits of adenosine triphosphate synthase in thermophilic bacterium PS3Biochimica et Biophysica Acta (BBA) - Bioenergetics, 1988
- The unusual enzymology of ATP synthaseBiochemistry, 1987
- Reconstitution of ATPase activity from the isolated α, β, and γ subunits of the coupling factor, F1, of Escherichia coliBiochemical and Biophysical Research Communications, 1977
- A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye bindingAnalytical Biochemistry, 1976
- The Mitochondrial ATPaseEuropean Journal of Biochemistry, 1975