Česká společnost pro hmotnostní spektrometrii

(13. ročník České konference hmotnostní spektrometrie a 11. Neformální proteomické setkání - WeO-05)
Insights into CyRepA2 primase-helicase-DNA interactions using structural mass spectrometry

Tomáš Smrčka ^1,2, Paulina Duhita Anindita ³, Jasmína Mária Portašíková ^1,2, Roman Sobotka ³, Alan Kádek ¹, Petr Man ^1 *

1. Mikrobiologický ústav AV ČR, v.v.i.
2. Přírodovědecká fakulta, Univerzita Karlova v Praze
3. Centrum Algatech, Mikrobiologický ústav AV ČR, v.v.i.

Abstrakt

Native mass spectrometry and H/D exchange are well-established structural mass spectrometry methods. Here we show how their combination could provide a complementary information of a dynamic protein-DNA interaction. Specifically, we examined the DNA binding of cyanobacterial primase-helicase CyRepA2.
Using native mass spectrometry, the oligomeric state and binding modes of CyRepA2 was examined in complex with several DNA ligands. With H/D exchange, the sites of DNA-induced structural changes were identified and a primase/helicase domain interface was probed. Furthermore, the role of ATP and AMP-PNP binding was also extensively studied providing insights into the dynamic nature of the enzyme’s function.
Overall, our work proves how the native mass spectrometry data together with the H/D exchange data nicely complement each other and together provide a comprehensive structural and functional insight into a highly dynamic protein complex.

* Korespondující autor: smrckat99@gmail.com