Tracking Ca2+ ATPase intermediates in real time by x-ray solution scattering

Matteo Levantino, Andreas Barth, Annette Duelli, Mattias Eklund, Michael Wulff, Chenge Li, Magnus Andersson, Harsha Ravishankar, Martin Nors Pedersen, Aljona Sitsel, Matteo Levantino, Claus Olesen, Poul Nissen

Risultato della ricerca: Articlepeer review

8 Citazioni (Scopus)

Abstract

Sarco/endoplasmic reticulum Ca2+ ATPase (SERCA) transporters regulate calcium signaling by active calcium ion reuptake to internal stores. Structural transitions associated with transport have been characterized by x-ray crystallography, but critical intermediates involved in the accessibility switch across the membrane are missing. We combined time-resolved x-ray solution scattering (TR-XSS) experiments and molecular dynamics (MD) simulations for real-time tracking of concerted SERCA reaction cycle dynamics in the native membrane. The equilibrium [Ca2] E1 state before laser activation differed in the domain arrangement compared with crystal structures, and following laser-induced release of caged ATP, a 1.5-ms intermediate was formed that showed closure of the cytoplasmic domains typical of E1 states with bound Ca2+ and ATP. A subsequent 13-ms transient state showed a previously unresolved actuator (A) domain arrangement that exposed the ADP-binding site after phosphorylation. Hence, the obtained TR-XSS models determine the relative timing of so-far elusive domain rearrangements in a native environment.
Lingua originaleEnglish
pagine (da-a)eaaz0981-
Numero di pagine11
RivistaScience advances
Volume6
Stato di pubblicazionePublished - 2020

All Science Journal Classification (ASJC) codes

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