The structure and function of P5A-ATPases.

Li, Ping; Bågenholm, Viktoria; Hägglund, Per; Lindkvist-Petersson, Karin; Wang, Kaituo; Gourdon, Pontus · Nat Commun · 2024

basic_science · Level V

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Abstract

Endoplasmic reticulum (ER) membrane resident P5A-ATPases broadly affect protein biogenesis and quality control, and yet their molecular function remains debated. Here, we report cryo-EM structures of a P5A-ATPase, CtSpf1, covering multiple transport intermediates of the E1 → E1-ATP → E1P-ADP → E1P → E2P → E2.P<sub>i</sub> → E2 → E1 cycle. In the E2P and E2.P<sub>i</sub> states a cleft spans the entire membrane, holding a polypeptide cargo molecule. The cargo includes an ER luminal extension, pinpointed as the C-terminus in the E2.P<sub>i</sub> state, which reenters the membrane in E2P. The E1 structure harbors a cytosol-facing cavity that is blocked by an insertion we refer to as the Plug-domain. The Plug-domain is nestled to key ATPase features and is displaced in the E1P-ADP and E1P states. Collectively, our findings are compatible with a broad range of proteins as cargo, with the P5A-ATPases serving a role in membrane removal of helices, although insertion/secretion cannot be excluded, as well as with a mechanistic role of the Plug-domain.

Medical subject headings