The Crystal Structure of the Ca2+-ATPase 1 from Listeria monocytogenes reveals a Pump Primed for Dephosphorylation

Sara Basse Hansen; Mateusz Dyla; Caroline Neumann; Esben Meldgaard Hoegh Quistgaard; Jacob Lauwring Andersen; Magnus Kjaergaard; Poul Nissen

doi:10.1016/j.jmb.2021.167015

The Crystal Structure of the Ca²⁺-ATPase 1 from Listeria monocytogenes reveals a Pump Primed for Dephosphorylation

J Mol Biol. 2021 Aug 6;433(16):167015. doi: 10.1016/j.jmb.2021.167015. Epub 2021 Apr 30.

Authors

Sara Basse Hansen¹, Mateusz Dyla¹, Caroline Neumann¹, Esben Meldgaard Hoegh Quistgaard¹, Jacob Lauwring Andersen¹, Magnus Kjaergaard², Poul Nissen³

Affiliations

¹ Department of Molecular Biology and Genetics, Aarhus University, Denmark; The Danish Research Institute for Translational Neuroscience (DANDRITE), Nordic EMBL Partnership for Molecular Medicine, Denmark.
² Department of Molecular Biology and Genetics, Aarhus University, Denmark; The Danish Research Institute for Translational Neuroscience (DANDRITE), Nordic EMBL Partnership for Molecular Medicine, Denmark; Aarhus Institute of Advanced Studies (AIAS), Denmark; The Danish National Research Foundation Center for Proteins in Memory (PROMEMO), Denmark.
³ Department of Molecular Biology and Genetics, Aarhus University, Denmark; The Danish Research Institute for Translational Neuroscience (DANDRITE), Nordic EMBL Partnership for Molecular Medicine, Denmark; The Danish National Research Foundation Center for Proteins in Memory (PROMEMO), Denmark. Electronic address: [email protected].

PMID: 33933469
DOI: 10.1016/j.jmb.2021.167015

Abstract

Many bacteria export intracellular calcium using active transporters homologous to the sarco/endoplasmic reticulum Ca²⁺-ATPase (SERCA). Here we present three crystal structures of Ca²⁺-ATPase 1 from Listeria monocytogenes (LMCA1). Structures with BeF₃^- mimicking a phosphoenzyme state reveal a closed state, which is intermediate between the outward-open E2P and the proton-occluded E2-P* conformations known for SERCA. It suggests that LMCA1 in the E2P state is pre-organized for dephosphorylation upon Ca²⁺ release, consistent with the rapid dephosphorylation observed in single-molecule studies. An arginine side-chain occupies the position equivalent to calcium binding site I in SERCA, leaving a single Ca²⁺ binding site in LMCA1, corresponding to SERCA site II. Observing no putative transport pathways dedicated to protons, we infer a direct proton counter transport through the Ca²⁺ exchange pathways. The LMCA1 structures provide insight into the evolutionary divergence and conserved features of this important class of ion transporters.

Keywords: Ca(2+)-ATPase LMCA1; Listeria; P-type ATPase; calcium; membrane protein crystallography.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Binding Sites*
Calcium / chemistry
Calcium / metabolism
Crystallography, X-Ray
Listeria monocytogenes / enzymology*
Models, Molecular*
Phosphorylation
Protein Binding*
Protein Conformation*
Sarcoplasmic Reticulum Calcium-Transporting ATPases / chemistry*
Sarcoplasmic Reticulum Calcium-Transporting ATPases / metabolism*
Structure-Activity Relationship*

Substances

Sarcoplasmic Reticulum Calcium-Transporting ATPases
Calcium