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

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

doi:10.1126/sciadv.aaz0981

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

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

^*Corresponding author af dette arbejde

Publikation: Bidrag til tidsskrift/Konferencebidrag i tidsskrift /Bidrag til avis › Tidsskriftartikel › Forskning › peer review

34 Citationer (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.

Originalsprog	Engelsk
Artikelnummer	eaaz0981
Tidsskrift	Science Advances
Vol/bind	6
Nummer	12
Antal sider	10
ISSN	2375-2548
DOI	https://doi.org/10.1126/sciadv.aaz0981
Status	Udgivet - mar. 2020

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10.1126/sciadv.aaz0981Licens: CC BY-NC

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http://www.scopus.com/inward/record.url?scp=85082174090&partnerID=8YFLogxK

Citationsformater

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title = "Tracking Ca2+ ATPase intermediates in real time by x-ray solution scattering",

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.",

keywords = "CA2+-ATPASE, CALCIUM-TRANSPORT, CONFORMATIONAL-CHANGES, MEMBRANE, MOLECULAR-DYNAMICS, PHOSPHOENZYME, PUMP, SARCOPLASMIC-RETICULUM, SKELETAL-MUSCLE, STRUCTURAL DYNAMICS",

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

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doi = "10.1126/sciadv.aaz0981",

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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 - 2020/3

Y1 - 2020/3

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.

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KW - MOLECULAR-DYNAMICS

KW - PHOSPHOENZYME

KW - PUMP

KW - SARCOPLASMIC-RETICULUM

KW - SKELETAL-MUSCLE

KW - STRUCTURAL DYNAMICS

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DO - 10.1126/sciadv.aaz0981

M3 - Journal article

C2 - 32219166

AN - SCOPUS:85082174090

SN - 2375-2548

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