Molecular Insights into Lipid-assisted Ca Regulation of the TRP Channel Polycystin-2

Overview

Journal Nat Struct Mol Biol

Specialties Cell Biology
Molecular Biology

Date 2017 Jan 17

PMID 28092368

Citations 68

Authors

Martin Wilkes

M Gregor Madej

Lydia Kreuter

Daniel Rhinow

Veronika Heinz

Silvia De Sanctis

Sabine Ruppel

Rebecca M Richter

Friederike Joos

Marina Grieben

Ashley C W Pike

Juha T Huiskonen

Elisabeth P Carpenter

Werner Kuhlbrandt

Ralph Witzgall

Christine Ziegler

Affiliations

Soon will be listed here.

Abstract

Polycystin-2 (PC2), a calcium-activated cation TRP channel, is involved in diverse Ca signaling pathways. Malfunctioning Ca regulation in PC2 causes autosomal-dominant polycystic kidney disease. Here we report two cryo-EM structures of distinct channel states of full-length human PC2 in complex with lipids and cations. The structures reveal conformational differences in the selectivity filter and in the large exoplasmic domain (TOP domain), which displays differing N-glycosylation. The more open structure has one cation bound below the selectivity filter (single-ion mode, PC2), whereas multiple cations are bound along the translocation pathway in the second structure (multi-ion mode, PC2). Ca binding at the entrance of the selectivity filter suggests Ca blockage in PC2, and we observed density for the Ca-sensing C-terminal EF hand in the unblocked PC2 state. The states show altered interactions of lipids with the pore loop and TOP domain, thus reflecting the functional diversity of PC2 at different locations, owing to different membrane compositions.

Citing Articles

A synthetic method to assay polycystin channel biophysics.

Larmore M, Esarte Palomero O, Kamat N, DeCaen P Elife. 2024; 13.

PMID: 39466685 PMC: 11517255. DOI: 10.7554/eLife.98534.

PKD2: An Important Membrane Protein in Organ Development.

Wang S, Kang Y, Xie H Cells. 2024; 13(20.

PMID: 39451240 PMC: 11506562. DOI: 10.3390/cells13201722.

Spontaneous Coronary Artery Dissection in Patients with Autosomal Dominant Polycystic Kidney Disease: A Systematic Review of the Literature.

Milutinovic S, Bell A, Jancic P, Stanojevic D, Borghol A, Mina J J Pers Med. 2024; 14(7).

PMID: 39063956 PMC: 11278354. DOI: 10.3390/jpm14070702.

Disease-associated missense mutations in the pore loop of polycystin-2 alter its ion channel function in a heterologous expression system.

Staudner T, Geiges L, Khamseekaew J, Sure F, Korbmacher C, Ilyaskin A J Biol Chem. 2024; 300(8):107574.

PMID: 39009345 PMC: 11630642. DOI: 10.1016/j.jbc.2024.107574.

A synthetic method to assay polycystin channel biophysics.

Larmore M, Esarte Palomero O, Kamat N, DeCaen P bioRxiv. 2024; .

PMID: 38766162 PMC: 11100589. DOI: 10.1101/2024.05.06.592666.

References

Kim S, Nie H, Nesin V, Tran U, Outeda P, Bai C . The polycystin complex mediates Wnt/Ca(2+) signalling. Nat Cell Biol. 2016; 18(7):752-764. PMC: 4925210. DOI: 10.1038/ncb3363. View

Gao Y, Cao E, Julius D, Cheng Y . TRPV1 structures in nanodiscs reveal mechanisms of ligand and lipid action. Nature. 2016; 534(7607):347-51. PMC: 4911334. DOI: 10.1038/nature17964. View

Mindell J, Grigorieff N . Accurate determination of local defocus and specimen tilt in electron microscopy. J Struct Biol. 2003; 142(3):334-47. DOI: 10.1016/s1047-8477(03)00069-8. View

Vassilev P, Guo L, Chen X, Segal Y, Peng J, Basora N . Polycystin-2 is a novel cation channel implicated in defective intracellular Ca(2+) homeostasis in polycystic kidney disease. Biochem Biophys Res Commun. 2001; 282(1):341-50. DOI: 10.1006/bbrc.2001.4554. View

Lyskov S, Chou F, Conchuir S, Der B, Drew K, Kuroda D . Serverification of molecular modeling applications: the Rosetta Online Server that Includes Everyone (ROSIE). PLoS One. 2013; 8(5):e63906. PMC: 3661552. DOI: 10.1371/journal.pone.0063906. View

Yu Y, Ulbrich M, Li M, Buraei Z, Chen X, Ong A . Structural and molecular basis of the assembly of the TRPP2/PKD1 complex. Proc Natl Acad Sci U S A. 2009; 106(28):11558-63. PMC: 2710685. DOI: 10.1073/pnas.0903684106. View

Bai X, Fernandez I, McMullan G, Scheres S . Ribosome structures to near-atomic resolution from thirty thousand cryo-EM particles. Elife. 2013; 2:e00461. PMC: 3576727. DOI: 10.7554/eLife.00461. View

Scheres S . Beam-induced motion correction for sub-megadalton cryo-EM particles. Elife. 2014; 3:e03665. PMC: 4130160. DOI: 10.7554/eLife.03665. View

Gallagher A, Hoffmann S, Brown N, Cedzich A, Meruvu S, Podlich D . A truncated polycystin-2 protein causes polycystic kidney disease and retinal degeneration in transgenic rats. J Am Soc Nephrol. 2006; 17(10):2719-30. DOI: 10.1681/ASN.2005090979. View

10.

Stamm M, Staritzbichler R, Khafizov K, Forrest L . Alignment of helical membrane protein sequences using AlignMe. PLoS One. 2013; 8(3):e57731. PMC: 3587630. DOI: 10.1371/journal.pone.0057731. View

11.

Scheres S . RELION: implementation of a Bayesian approach to cryo-EM structure determination. J Struct Biol. 2012; 180(3):519-30. PMC: 3690530. DOI: 10.1016/j.jsb.2012.09.006. View

12.

Yang Y, Ehrlich B . Structural studies of the C-terminal tail of polycystin-2 (PC2) reveal insights into the mechanisms used for the functional regulation of PC2. J Physiol. 2016; 594(15):4141-9. PMC: 4967733. DOI: 10.1113/JP270933. View

13.

Afonine P, Grosse-Kunstleve R, Echols N, Headd J, Moriarty N, Mustyakimov M . Towards automated crystallographic structure refinement with phenix.refine. Acta Crystallogr D Biol Crystallogr. 2012; 68(Pt 4):352-67. PMC: 3322595. DOI: 10.1107/S0907444912001308. View

14.

Narayan K, Subramaniam S . Focused ion beams in biology. Nat Methods. 2015; 12(11):1021-31. PMC: 6993138. DOI: 10.1038/nmeth.3623. View

15.

Kottgen M, Buchholz B, Garcia-Gonzalez M, Kotsis F, Fu X, Doerken M . TRPP2 and TRPV4 form a polymodal sensory channel complex. J Cell Biol. 2008; 182(3):437-47. PMC: 2500130. DOI: 10.1083/jcb.200805124. View

16.

Li X, Mooney P, Zheng S, Booth C, Braunfeld M, Gubbens S . Electron counting and beam-induced motion correction enable near-atomic-resolution single-particle cryo-EM. Nat Methods. 2013; 10(6):584-90. PMC: 3684049. DOI: 10.1038/nmeth.2472. View

17.

Qian F, Germino F, Cai Y, Zhang X, Somlo S, Germino G . PKD1 interacts with PKD2 through a probable coiled-coil domain. Nat Genet. 1997; 16(2):179-83. DOI: 10.1038/ng0697-179. View

18.

Song Y, DiMaio F, Wang R, Kim D, Miles C, Brunette T . High-resolution comparative modeling with RosettaCM. Structure. 2013; 21(10):1735-42. PMC: 3811137. DOI: 10.1016/j.str.2013.08.005. View

19.

Grant T, Grigorieff N . Measuring the optimal exposure for single particle cryo-EM using a 2.6 Å reconstruction of rotavirus VP6. Elife. 2015; 4:e06980. PMC: 4471936. DOI: 10.7554/eLife.06980. View

20.

Tsiokas L, Kim E, Arnould T, Sukhatme V, Walz G . Homo- and heterodimeric interactions between the gene products of PKD1 and PKD2. Proc Natl Acad Sci U S A. 1997; 94(13):6965-70. PMC: 21268. DOI: 10.1073/pnas.94.13.6965. View