Premium
In vivo affinity label of a protein expressed in Escherichia coli
Author(s) -
Odaka Masafumi,
Kiribuchi Kyoko,
Allison William S.,
Yoshida Masasuke
Publication year - 1993
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 - 10.1016/0014-5793(93)80809-9
Subject(s) - protein subunit , escherichia coli , cysteine , mutant , biochemistry , population , chemistry , cofactor , biology , microbiology and biotechnology , enzyme , gene , demography , sociology
When Tyr‐307 of the β subunit of f 1 ‐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 α 3 β 3 , 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.