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Pounot Kevin

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Lukács András
Conformational flexibility in a photoactivated adenylate cyclase studied by fluorescence spectroscopy and solution X-ray scattering

Aug 29 - kedd

09:10 – 09:30

Molekuláris biofizika

E03

Conformational flexibility in a photoactivated adenylate cyclase studied by fluorescence spectroscopy and solution X-ray scattering

1,2, , , András Kengyel, Matteo Levantino, Caroline Maas, Dihia Moussaoui, Ildikó Pécsi, Petra Pernot, Kevin Pounot, Giorgio Schiro, Mark Tully, Kinga Ujfalusi-Pozsonyi, Jovana Vitas, Martin Weik, András Lukács

Sofia Maria Kapetanaki^1,2, Emőke Bódis¹, Miklós Nyitrai¹, András Kengyel¹, Matteo Levantino³, Caroline Maas², Dihia Moussaoui³, Ildikó Pécsi¹, Petra Pernot³, Kevin Pounot^2,3, Giorgio Schirò², Mark Tully³, Kinga Ujfalusi-Pozsonyi¹, Jovana Vitas², Martin Weik², András Lukács¹

¹ Department of Biophysics, Medical School, University of Pécs, 7624 Pécs, Hungary

² CEA–Institut de Biologie Structurale, Grenoble, 38044 France

³ European Synchrotron Radiation Facility, Grenoble, 38043 France

The photoactivated adenylate cyclase from the photosynthetic cyanobacterium Oscillatoria acuminata OaPAC is a homodimeric enzyme comprising of a N-terminal domain that senses blue light using flavin (BLUF) [1] and a C-terminal class III adenylate cyclase (AC) domain that catalyses the formation of cAMP from ATP (adenosine triphosphate) [2,3]. cAMP is a universal regulator of metabolism and gene expression in all life forms [4]. Modulating the cellular concentration of cAMP has emerged in the focus of modern optogenetic applications and therapeutic approaches. Recent crystallographic studies have indicated that the activation mechanism of OaPAC involves only small movements. In this study [5], we apply small-angle X-ray scattering (SAXS) [6] and time-resolved solution X-ray scattering [7] in combination with other biophysical techniques to investigate the substrate induced-conformational changes of OaPAC in solution. The implications of our work to the function of the enzyme are discussed.

Acknowledgment

A.L. acknowledges funding from the Hungarian National Research and Innovation Office (K-137557) and was supported by PTE ÁOK-KA-2021

References

[1] Fujisawa, T. and Masuda, S. (2018) Light-induced chromophore and protein responses and mechanical signal transduction of BLUF proteins Biophys. Rev. 10, 327-337.

[2] Ohki, M. et al. (2016) Structural insight into photoactivation of an adenylate cyclase from a photosynthetic cyanobacterium Proc. Natl. Acad. Sci. 113, 6659-6664.

[3] Ohki, M. et al. (2017) Molecular mechanism of photoactivation of a light-regulated adenylate cyclase Proc. Natl. Acad. Sci. 114, 8562-8567.

[4] Zaccolo, M., Zerio, A., and Lobo, M.J. (2021) Subcellular Organization of the cAMP Signaling Pathway Pharmacol.Rev. 73, 278–309.

[5] Kapetanaki, S.M. et al. (unpublished results)

[6] Da Vela Stefano and Svergun, D. (2020) Methods, development and applications of small-angle X-ray scattering to characterize biological macromolecules in solution Curr. Res. Struct. Biol. 2, 164-170.

[7] Cho, H.S., Schotte, F., Stadnytskyi, V., and Anfinrud, P. (2021) Time-resolved X-ray scattering studies of proteins Curr. Opin. Struct. Biol. 70, 99-107.