Kropp, Cosimo and Bruckmann, Astrid and Babinger, Patrick (2022) Controlling Enzymatic Activity by Modulating the Oligomerization State via Chemical Rescue and Optical Control. CHEMBIOCHEM, 23 (5). ISSN 1439-4227, 1439-7633
Full text not available from this repository. (Request a copy)Abstract
Selective switching of enzymatic activity has been a longstanding goal in synthetic biology. Drastic changes in activity upon mutational manipulation of the oligomerization state of enzymes have frequently been reported in the literature, but scarcely exploited for switching. Using geranylgeranylglyceryl phosphate synthase as a model, we demonstrate that catalytic activity can be efficiently controlled by exogenous modulation of the association state. We introduced a lysine-to-cysteine mutation, leading to the breakdown of the active hexamer into dimers with impaired catalytic efficiency. Addition of bromoethylamine chemically rescued the enzyme by restoring hexamerization and activity. As an alternative method, we incorporated the photosensitive unnatural amino acid o-nitrobenzyl-O-tyrosine (ONBY) into the hexamerization interface. This again led to inactive dimers, but the hexameric state and activity could be recovered by UV-light induced cleavage of ONBY. For both approaches, we obtained switching factors greater than 350-fold, which compares favorably with previously reported activity changes that were caused by site-directed mutagenesis.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | BRONSTED ANALYSIS; PHOSPHATE; SPECIFICITY; MECHANISMS; EVOLUTION; MUTANT; biocatalysis; chemical rescue; oligomerization; optochemical tools; protein-protein interactions |
| Subjects: | 500 Science > 570 Life sciences |
| Divisions: | Biology, Preclinical Medicine > Institut für Biophysik und physikalische Biochemie Biology, Preclinical Medicine > Institut für Biochemie, Genetik und Mikrobiologie > Lehrstuhl für Biochemie I > Prof. Dr. Gunter Meister |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 05 Jul 2022 14:25 |
| Last Modified: | 05 Jul 2022 14:25 |
| URI: | https://pred.uni-regensburg.de/id/eprint/45608 |
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