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Structure of the [Ca]E2P intermediate of Ca2+-ATPase 1 from Listeria monocytogenes

ABSTRACT: Active transport by P-type Ca2+-ATPases maintain internal calcium stores and a low cytosolic calcium concentration. Structural studies of mammalian sarco/endoplasmic reticulum Ca2+-ATPases (SERCA) have revealed several steps of the transport cycle, but a calcium releasing intermediate has remained elusive. Single-molecule FRET studies of the bacterial Ca2+-ATPase LMCA1 revealed an intermediate of the transition between so-called [Ca]E1P and E2P states and suggested that calcium release from this intermediate was the essentially irreversible step of transport. Here, we present a 3.5 Å resolution cryo-EM structure for a four-glycine insertion mutant of LMCA1 in a lipid nanodisc obtained under conditions with calcium and ATP and adopting such an intermediate state, denoted [Ca]E2P. The cytosolic domains are positioned in the E2P-like conformation, while the calcium-binding transmembrane (TM) domain adopts a calcium-bound E1P-ADP like conformation. Missing density for the E292 residue at the calcium site (equivalent of SERCA1a E309) suggests flexibility and a site poised for calcium release and proton uptake. The structure suggests a mechanism where ADP release and re-organisation of the cytoplasmic domains precede calcium release

SUBMITTER: Poul Nissen

PROVIDER: S-SCDT-10_1038-S44319-025-00392-X | biostudies-other |

REPOSITORIES: biostudies-other

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