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

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.