Single amino acid mutation decouples photochemistry of the BLUF domain from the enzymatic function of OaPAC and drives the enzyme to a switched-on state

Tolentino Collado, Jinnette, Bodis, Emoke, Pasitka, Jonatan, Szucs, Mihaly, Fekete, Zsuzsanna, Kis-Bicskei, Nikolett, Telek, Elek, Pozsonyi, Kinga, Kapetanaki, Sofia M., Greetham, Greg, Tonge, Peter J., Meech, Stephen R. ORCID: https://orcid.org/0000-0001-5561-2782 and Lukacs, Andras (2023) Single amino acid mutation decouples photochemistry of the BLUF domain from the enzymatic function of OaPAC and drives the enzyme to a switched-on state. Journal of Molecular Biology. ISSN 0022-2836

Full text not available from this repository. (Request a copy)

Abstract

Photoactivated adenylate cyclases (PACs) are light-activated enzymes that combine a BLUF (blue-light using flavin) domain and an adenylate cyclase domain that are able to increase the levels of the important second messenger cAMP (cyclic adenosine monophosphate) upon blue-light excitation. The light-induced changes in the BLUF domain are transduced to the adenylate cyclase domain via a mechanism that has not yet been established. One critical residue in the photoactivation mechanism of BLUF domains, present in the vicinity of the flavin is the glutamine amino acid close to the N5 of the flavin. The role of this residue has been investigated extensively both experimentally and theoretically. However, its role in the activity of the photoactivated adenylate cyclase, OaPAC has never been addressed. In this work, we applied ultrafast transient visible and infrared spectroscopies to study the photochemistry of the Q48E OaPAC mutant. This mutation altered the primary electron transfer process and switched the enzyme into a permanent ‘on’ state, able to increase the cAMP levels under dark conditions compared to the cAMP levels of the dark-adapted state of the wild-type OaPAC. Differential scanning calorimetry measurements point to a less compact structure for the Q48E OaPAC mutant. The ensemble of these findings provide insight into the important elements in PACs and how their fine tuning may help in the design of optogenetic devices.

Item Type: Article
Faculty \ School: Faculty of Science > School of Chemistry
UEA Research Groups: Faculty of Science > Research Groups > Centre for Photonics and Quantum Science
Faculty of Science > Research Groups > Chemistry of Light and Energy
Depositing User: LivePure Connector
Date Deposited: 17 Oct 2023 00:45
Last Modified: 17 Oct 2023 00:46
URI: https://ueaeprints.uea.ac.uk/id/eprint/93323
DOI: 10.1016/j.jmb.2023.168312

Actions (login required)

View Item View Item