TY - JOUR
T1 - The SERCA residue Glu340 mediates interdomain communication that guides Ca2+transport
AU - Geurts, Maxwell M.G.
AU - Clausen, Johannes D.
AU - Arnou, Bertrand
AU - Montigny, Cédric
AU - Lenoir, Guillaume
AU - Corey, Robin A.
AU - Jaxel, Christine
AU - Møller, Jesper V.
AU - Nissen, Poul
AU - Andersen, Jens Peter
AU - Le Maire, Marc
AU - Bublitz, Maike
N1 - Funding Information:
ACKNOWLEDGMENTS. This work was supported by the French Infrastructure for Integrated Structural Biology (ANR-10-INSB-05), and the CNRS, the Agence Nationale de la Recherche and the Domaines d’Intérêt Majeur Maladies Infectieuses Région Ile de-France (to M.l.M., C.J., G.L., and C.M.), the Center for Membrane Pumps in Cells and Disease of the Danish National Research Foundation (to J.V.M., P.N., J.P.A., and M.B.), the Lundbeck Foundation (to P.N.), the Wellcome Trust (ref. 220063/Z/20/Z to M.M.G.G.), and the Danish Council for Independent Research (to J.P.A.). We are grateful to Anna Marie Nielsen for technical support and Jesper L. Karlsen and Mark Sansom for guidance and support regarding scientific computing as well as access to facilities.
Funding Information:
This work was supported by the French Infrastructure for Integrated Structural Biology (ANR-10- INSB-05), and the CNRS, the Agence Nationale de la Recherche and the Domaines d'Int?r?t Majeur Maladies Infectieuses R?gion Ile de-France (to M.l.M., C.J., G.L., and C.M.), the Center for Membrane Pumps in Cells and Disease of the Danish National Research Foundation (to J.V.M., P.N., J.P.A., and M.B.), the Lundbeck Foundation (to P.N.), the Wellcome Trust (ref. 220063/Z/20/Z to M.M.G.G.), and the Danish Council for Independent Research (to J.P.A.). We are grateful to Anna Marie Nielsen for technical support and Jesper L. Karlsen and Mark Sansom for guidance and support regarding scientific computing as well as access to facilities.
Publisher Copyright:
© 2020 National Academy of Sciences. All rights reserved.
Copyright:
Copyright 2020 Elsevier B.V., All rights reserved.
PY - 2020/12
Y1 - 2020/12
N2 - The sarco(endo)plasmic reticulum Ca2+-ATPase (SERCA) is a P-type ATPase that transports Ca2+ from the cytosol into the sarco(endo) plasmic reticulum (SR/ER) lumen, driven by ATP. This primary transport activity depends on tight coupling between movements of the transmembrane helices forming the two Ca2+-binding sites and the cytosolic headpiece mediating ATP hydrolysis. We have addressed the molecular basis for this intramolecular communication by analyzing the structure and functional properties of the SERCA mutant E340A. The mutated Glu340 residue is strictly conserved among the P-type ATPase family of membrane transporters and is located at a seemingly strategic position at the interface between the phosphorylation domain and the cytosolic ends of 5 of SERCA's 10 transmembrane helices. The mutant displays a marked slowing of the Ca2+-binding kinetics, and its crystal structure in the presence of Ca2+ and ATP analog reveals a rotated headpiece, altered connectivity between the cytosolic domains, and an altered hydrogen bonding pattern around residue 340. Supported by molecular dynamics simulations, we conclude that the E340A mutation causes a stabilization of the Ca2+ sites in a more occluded state, hence displaying slowed dynamics. This finding underpins a crucial role of Glu340 in interdomain communication between the headpiece and the Ca2+-binding transmembrane region.
AB - The sarco(endo)plasmic reticulum Ca2+-ATPase (SERCA) is a P-type ATPase that transports Ca2+ from the cytosol into the sarco(endo) plasmic reticulum (SR/ER) lumen, driven by ATP. This primary transport activity depends on tight coupling between movements of the transmembrane helices forming the two Ca2+-binding sites and the cytosolic headpiece mediating ATP hydrolysis. We have addressed the molecular basis for this intramolecular communication by analyzing the structure and functional properties of the SERCA mutant E340A. The mutated Glu340 residue is strictly conserved among the P-type ATPase family of membrane transporters and is located at a seemingly strategic position at the interface between the phosphorylation domain and the cytosolic ends of 5 of SERCA's 10 transmembrane helices. The mutant displays a marked slowing of the Ca2+-binding kinetics, and its crystal structure in the presence of Ca2+ and ATP analog reveals a rotated headpiece, altered connectivity between the cytosolic domains, and an altered hydrogen bonding pattern around residue 340. Supported by molecular dynamics simulations, we conclude that the E340A mutation causes a stabilization of the Ca2+ sites in a more occluded state, hence displaying slowed dynamics. This finding underpins a crucial role of Glu340 in interdomain communication between the headpiece and the Ca2+-binding transmembrane region.
KW - Cabinding
KW - Molecular dynamics simulations
KW - P-type ATPase
KW - SERCA
KW - Tryptophan fluorescence
UR - http://www.scopus.com/inward/record.url?scp=85097581125&partnerID=8YFLogxK
U2 - 10.1073/pnas.2014896117
DO - 10.1073/pnas.2014896117
M3 - Journal article
C2 - 33229570
AN - SCOPUS:85097581125
SN - 0027-8424
VL - 117
SP - 31114
EP - 31122
JO - Proceedings of the National Academy of Sciences (PNAS)
JF - Proceedings of the National Academy of Sciences (PNAS)
IS - 49
ER -