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Optical Control of Cellular ATP Levels with a Photocaged Adenylate Kinase
Author(s) -
Zhou Wenyuan,
Hankinson Chasity P.,
Deiters Alexander
Publication year - 2020
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.201900757
Subject(s) - adenylate kinase , biochemistry , chemistry , lysine , mutagenesis , microbiology and biotechnology , enzyme , biology , mutant , amino acid , gene
We have developed a new tool for the optical control of cellular ATP concentrations with a photocaged adenylate kinase (Adk). The photocaged Adk is generated by substituting a catalytically essential lysine with a hydroxycoumarin‐protected lysine through site‐specific unnatural amino acid mutagenesis in both E. coli and mammalian cells. Caging of the critical lysine residue offers complete suppression of Adk's phosphotransferase activity and rapid restoration of its function both in vitro and in vivo upon optical stimulation. Light‐activated Adk renders faster rescue of cell growth than chemically inducible expression of wild‐type Adk in E. coli as well as rapid ATP depletion in mammalian cells. Thus, caging Adk provides a new tool for direct conditional perturbation of cellular ATP concentrations thereby enabling the investigation of ATP‐coupled physiological events in temporally dynamic contexts.