SERCA mutant E309Q binds two Ca ions but adopts a catalytically incompetent conformation

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Standard

SERCA mutant E309Q binds two Ca ions but adopts a catalytically incompetent conformation. / Clausen, Johannes D.; Bublitz, Maike; Arnou, Bertrand; Montigny, Cédric; Jaxel, Christine; Møller, Jesper Vuust; Nissen, Poul; Andersen, Jens Peter; le Maire, Marc.

In: E M B O Journal, Vol. 32, No. 24, 11.12.2013, p. 3231-3243.

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaperJournal articleResearchpeer-review

Harvard

Clausen, JD, Bublitz, M, Arnou, B, Montigny, C, Jaxel, C, Møller, JV, Nissen, P, Andersen, JP & le Maire, M 2013, 'SERCA mutant E309Q binds two Ca ions but adopts a catalytically incompetent conformation', E M B O Journal, vol. 32, no. 24, pp. 3231-3243. https://doi.org/10.1038/emboj.2013.250

APA

Clausen, J. D., Bublitz, M., Arnou, B., Montigny, C., Jaxel, C., Møller, J. V., ... le Maire, M. (2013). SERCA mutant E309Q binds two Ca ions but adopts a catalytically incompetent conformation. E M B O Journal, 32(24), 3231-3243. https://doi.org/10.1038/emboj.2013.250

CBE

Clausen JD, Bublitz M, Arnou B, Montigny C, Jaxel C, Møller JV, Nissen P, Andersen JP, le Maire M. 2013. SERCA mutant E309Q binds two Ca ions but adopts a catalytically incompetent conformation. E M B O Journal. 32(24):3231-3243. https://doi.org/10.1038/emboj.2013.250

MLA

Vancouver

Clausen JD, Bublitz M, Arnou B, Montigny C, Jaxel C, Møller JV et al. SERCA mutant E309Q binds two Ca ions but adopts a catalytically incompetent conformation. E M B O Journal. 2013 Dec 11;32(24):3231-3243. https://doi.org/10.1038/emboj.2013.250

Author

Clausen, Johannes D. ; Bublitz, Maike ; Arnou, Bertrand ; Montigny, Cédric ; Jaxel, Christine ; Møller, Jesper Vuust ; Nissen, Poul ; Andersen, Jens Peter ; le Maire, Marc. / SERCA mutant E309Q binds two Ca ions but adopts a catalytically incompetent conformation. In: E M B O Journal. 2013 ; Vol. 32, No. 24. pp. 3231-3243.

Bibtex

@article{6c451a5eb47848d4a3f9e59dcb99b715,
title = "SERCA mutant E309Q binds two Ca ions but adopts a catalytically incompetent conformation",
abstract = "The sarco(endo)plasmic reticulum Ca2+-ATPase (SERCA) couples ATP hydrolysis to transport of Ca2+. This directed energy transfer requires cross-talk between the two Ca2+ sites and the phosphorylation site over 50 {\AA} distance. We have addressed the mechano-structural basis for this intramolecular signal by analysing the structure and the functional properties of SERCA mutant E309Q. Glu309 contributes to Ca2+ coordination at site II, and a consensus has been that E309Q only binds Ca2+ at site I. The crystal structure of E309Q in the presence of Ca2+ and an ATP analogue, however, reveals two occupied Ca2+ sites of a non-catalytic Ca2E1 state. Ca2+ is bound with micromolar affinity by both Ca2+ sites in E309Q, but without cooperativity. The Ca2+-bound mutant does phosphorylate from ATP, but at a very low maximal rate. Phosphorylation depends on the correct positioning of the A-domain, requiring a shift of transmembrane segment M1 into an ‘up and kinked position’. This transition is impaired in the E309Q mutant, most likely due to a lack of charge neutralization and altered hydrogen binding capacities at Ca2+ site II.",
author = "Clausen, {Johannes D.} and Maike Bublitz and Bertrand Arnou and C{\'e}dric Montigny and Christine Jaxel and M{\o}ller, {Jesper Vuust} and Poul Nissen and Andersen, {Jens Peter} and {le Maire}, Marc",
year = "2013",
month = "12",
day = "11",
doi = "10.1038/emboj.2013.250",
language = "English",
volume = "32",
pages = "3231--3243",
journal = "E M B O Journal",
issn = "0261-4189",
publisher = "Wiley-Blackwell Publishing Ltd",
number = "24",

}

RIS

TY - JOUR

T1 - SERCA mutant E309Q binds two Ca ions but adopts a catalytically incompetent conformation

AU - Clausen, Johannes D.

AU - Bublitz, Maike

AU - Arnou, Bertrand

AU - Montigny, Cédric

AU - Jaxel, Christine

AU - Møller, Jesper Vuust

AU - Nissen, Poul

AU - Andersen, Jens Peter

AU - le Maire, Marc

PY - 2013/12/11

Y1 - 2013/12/11

N2 - The sarco(endo)plasmic reticulum Ca2+-ATPase (SERCA) couples ATP hydrolysis to transport of Ca2+. This directed energy transfer requires cross-talk between the two Ca2+ sites and the phosphorylation site over 50 Å distance. We have addressed the mechano-structural basis for this intramolecular signal by analysing the structure and the functional properties of SERCA mutant E309Q. Glu309 contributes to Ca2+ coordination at site II, and a consensus has been that E309Q only binds Ca2+ at site I. The crystal structure of E309Q in the presence of Ca2+ and an ATP analogue, however, reveals two occupied Ca2+ sites of a non-catalytic Ca2E1 state. Ca2+ is bound with micromolar affinity by both Ca2+ sites in E309Q, but without cooperativity. The Ca2+-bound mutant does phosphorylate from ATP, but at a very low maximal rate. Phosphorylation depends on the correct positioning of the A-domain, requiring a shift of transmembrane segment M1 into an ‘up and kinked position’. This transition is impaired in the E309Q mutant, most likely due to a lack of charge neutralization and altered hydrogen binding capacities at Ca2+ site II.

AB - The sarco(endo)plasmic reticulum Ca2+-ATPase (SERCA) couples ATP hydrolysis to transport of Ca2+. This directed energy transfer requires cross-talk between the two Ca2+ sites and the phosphorylation site over 50 Å distance. We have addressed the mechano-structural basis for this intramolecular signal by analysing the structure and the functional properties of SERCA mutant E309Q. Glu309 contributes to Ca2+ coordination at site II, and a consensus has been that E309Q only binds Ca2+ at site I. The crystal structure of E309Q in the presence of Ca2+ and an ATP analogue, however, reveals two occupied Ca2+ sites of a non-catalytic Ca2E1 state. Ca2+ is bound with micromolar affinity by both Ca2+ sites in E309Q, but without cooperativity. The Ca2+-bound mutant does phosphorylate from ATP, but at a very low maximal rate. Phosphorylation depends on the correct positioning of the A-domain, requiring a shift of transmembrane segment M1 into an ‘up and kinked position’. This transition is impaired in the E309Q mutant, most likely due to a lack of charge neutralization and altered hydrogen binding capacities at Ca2+ site II.

U2 - 10.1038/emboj.2013.250

DO - 10.1038/emboj.2013.250

M3 - Journal article

VL - 32

SP - 3231

EP - 3243

JO - E M B O Journal

JF - E M B O Journal

SN - 0261-4189

IS - 24

ER -