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TitleStructure of the [Ca]E2P intermediate of Ca-ATPase 1 from Listeria monocytogenes.
Journal, issue, pagesEMBO Rep, Vol. 26, Issue 7, Page 1709-1723, Year 2025
Publish dateFeb 27, 2025
AuthorsSara Basse Hansen / Rasmus Kock Flygaard / Magnus Kjaergaard / Poul Nissen /
PubMed AbstractActive transport by P-type Ca-ATPases maintain internal calcium stores and a low cytosolic calcium concentration. Structural studies of mammalian sarco/endoplasmic reticulum Ca-ATPases (SERCA) have ...Active transport by P-type Ca-ATPases maintain internal calcium stores and a low cytosolic calcium concentration. Structural studies of mammalian sarco/endoplasmic reticulum Ca-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 Ca-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 (the 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-organization of the cytoplasmic domains precede calcium release.
External linksEMBO Rep / PubMed:40016426 / PubMed Central
MethodsEM (single particle)
Resolution3.46 Å
Structure data

51510

9gqo

EMDB-51510, PDB-9gqo:
Structure of a Ca2+ bound phosphoenzyme intermediate in the inward-to-outward transition of Ca2+-ATPase 1 from Listeria monocytogenes
Method: EM (single particle) / Resolution: 3.46 Å

Chemicals

ChemComp-CA:
Unknown entry

ChemComp-MG:
Unknown entry

Source
  • listeria monocytogenes (bacteria)
KeywordsMEMBRANE PROTEIN / Intermediate / P-type ATPase / Calcium Transport

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