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A high‐throughput genetically directed protein crosslinking analysis reveals the physiological relevance of the ATP synthase ‘inserted’ state
Author(s) -
Liu Yang,
Yu Jiayu,
Wang Mengyuan,
Zeng Qingfang,
Fu Xinmiao,
Chang Zengyi
Publication year - 2021
Publication title -
the febs journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.981
H-Index - 204
eISSN - 1742-4658
pISSN - 1742-464X
DOI - 10.1111/febs.15616
Subject(s) - atp synthase , protein subunit , biochemistry , biology , mutagenesis , enzyme , atpase , chemistry , microbiology and biotechnology , gene , mutation
We developed a large scale in vivo protein photocrosslinking analysis pipeline based on the introduction of unnatural amino acid into target protein via scarless genome‐targeted site‐directed mutagenesis, and probing cross‐linked products via high‐throughput polyacrylamide gel electrophoresis. We uncovered that bacterial ATP synthase exists as an equilibrium between the ‘inserted’ and ‘noninserted’ state, maintaining a proper level of net ATP synthesis when shifting to the former under unfavorable energetically stressful conditions.