Tracking Ca2+ ATPase intermediates in real time by x-ray solution scattering
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Tracking Ca2+ ATPase intermediates in real time by x-ray solution scattering. / Ravishankar, Harsha; Pedersen, Martin Nors; Eklund, Mattias; Sitsel, Aljona; Li, Chenge; Duelli, Annette; Levantino, Matteo; Wulff, Michael; Barth, Andreas; Olesen, Claus; Nissen, Poul; Andersson, Magnus.
I: Science Advances, Bind 6, Nr. 12, eaaz0981, 2020.Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt
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TY - JOUR
T1 - Tracking Ca2+ ATPase intermediates in real time by x-ray solution scattering
AU - Ravishankar, Harsha
AU - Pedersen, Martin Nors
AU - Eklund, Mattias
AU - Sitsel, Aljona
AU - Li, Chenge
AU - Duelli, Annette
AU - Levantino, Matteo
AU - Wulff, Michael
AU - Barth, Andreas
AU - Olesen, Claus
AU - Nissen, Poul
AU - Andersson, Magnus
PY - 2020
Y1 - 2020
N2 - 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.
AB - 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.
U2 - 10.1126/sciadv.aaz0981
DO - 10.1126/sciadv.aaz0981
M3 - Journal article
C2 - 32219166
AN - SCOPUS:85082174090
VL - 6
JO - Science advances
JF - Science advances
SN - 2375-2548
IS - 12
M1 - eaaz0981
ER -
ID: 239812687