{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,15]],"date-time":"2026-10-15T05:29:32Z","timestamp":1792042172797,"version":"4.4.0"},"reference-count":65,"publisher":"National Academy of Sciences","issue":"2","license":[{"start":{"date-parts":[[2026,1,5]],"date-time":"2026-01-05T00:00:00Z","timestamp":1767571200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001809","name":"MOST | National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["32271261"],"award-info":[{"award-number":["32271261"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100017623","name":"MOST | NSFC | NSFC-Zhejiang Joint Fund | \u6d59\u6c5f\u7701\u79d1\u5b66\u6280\u672f\u5385 | ZJNSF | Zhejiang Provincial Outstanding Youth Science Foundation","doi-asserted-by":"publisher","award":["LR22C050003"],"award-info":[{"award-number":["LR22C050003"]}],"id":[{"id":"10.13039\/501100017623","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["www.pnas.org"],"crossmark-restriction":true},"short-container-title":["Proc. Natl. Acad. Sci. U.S.A."],"published-print":{"date-parts":[[2026,1,13]]},"abstract":"<jats:p>\n                    CatSper serves as the primary Ca\n                    <jats:sup>2+<\/jats:sup>\n                    entry pathway in the principal piece of sperm flagellum and is crucial for sperm motility and fertility. Sperm lacking functional CatSper channels fail to undergo hyperactivation during fertilization, leading to complete male infertility. Along the longitudinal axis of the sperm flagellum, staggered CatSper complexes align in a hand-in-hand arrangement, forming a distinctive quadrilinear zigzag arrays known as CatSper nanodomains. However, the molecular details of how CatSper oligomerizes to form such higher-order assembly remain unclear. In this study, we present the cryoelectron microscopy (cryo-EM) structures of native CatSper dimer (~1.5 MDa) and trimer (~2.3 MDa) megacomplexes, which represent the fundamental units of the zigzag array. We reveal the overall configuration of the zigzag assembly by characterizing the two distinct dimer interfaces that mediate CatSper oligomerization. Specifically, we elucidate the interaction details of two extracellular interfaces formed by CATSPER\u03b2 and CATSPER\u03b3 at the two dimer interfaces, respectively, and find that CATSPER\u03b7, a recently identified component of CatSper, constitutes the transmembrane interface within one of these dimer interfaces. Functional studies in mice demonstrate that CATSPER\u03b7 is essential for the formation of functional CatSper on the sperm flagellum, and CATSPER\u03b7-deficient sperm fail to hyperactivate during fertilization, resulting in male infertility both in vivo and in vitro. Our data provide insights into the higher-order assembly of CatSper at the molecular level, offering clues for the development of male contraceptives.\n                  <\/jats:p>","DOI":"10.1073\/pnas.2510754123","type":"journal-article","created":{"date-parts":[[2026,1,5]],"date-time":"2026-01-05T20:02:28Z","timestamp":1767643348000},"update-policy":"https:\/\/doi.org\/10.1073\/pnas.cm10313","source":"Crossref","is-referenced-by-count":4,"title":["Molecular basis of the higher-order assembly of CatSper"],"prefix":"10.1073","volume":"123","author":[{"given":"Qikui","family":"Xu","sequence":"first","affiliation":[{"name":"State Key Laboratory of Gene Expression, School of Life Sciences, Westlake University","place":["Hangzhou, Zhejiang, China"]},{"name":"Zhejiang Key Laboratory of Structural Biology, School of Life Sciences, Westlake University","place":["Hangzhou, China"]},{"name":"Westlake Laboratory of Life Sciences and Biomedicine","place":["Hangzhou, China"]},{"name":"Institute of Biology, Westlake Institute for Advanced Study","place":["Hangzhou, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9026-8275","authenticated-orcid":false,"given":"Shiyi","family":"Lin","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Gene Expression, School of Life Sciences, Westlake University","place":["Hangzhou, Zhejiang, China"]},{"name":"Zhejiang Key Laboratory of Structural Biology, School of Life Sciences, Westlake University","place":["Hangzhou, China"]},{"name":"Westlake Laboratory of Life Sciences and Biomedicine","place":["Hangzhou, China"]},{"name":"Institute of Biology, Westlake Institute for Advanced Study","place":["Hangzhou, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qingqing","family":"Zhao","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Gene Expression, School of Life Sciences, Westlake University","place":["Hangzhou, Zhejiang, China"]},{"name":"Zhejiang Key Laboratory of Structural Biology, School of Life Sciences, Westlake University","place":["Hangzhou, China"]},{"name":"Westlake Laboratory of Life Sciences and Biomedicine","place":["Hangzhou, China"]},{"name":"Institute of Biology, Westlake Institute for Advanced Study","place":["Hangzhou, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yanfei","family":"Ru","sequence":"additional","affiliation":[{"name":"Westlake Laboratory of Life Sciences and Biomedicine","place":["Hangzhou, China"]},{"name":"Institute of Biology, Westlake Institute for Advanced Study","place":["Hangzhou, China"]},{"name":"Key Laboratory of Growth Regulation and Translational Research of Zhejiang Province, School of Life Sciences, Westlake University","place":["Hangzhou, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hang","family":"Kang","sequence":"additional","affiliation":[{"name":"Institute of Reproductive Medicine, Medical school, Nantong University","place":["Nantong, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xuhui","family":"Zeng","sequence":"additional","affiliation":[{"name":"Institute of Reproductive Medicine, Medical school, Nantong University","place":["Nantong, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7005-937X","authenticated-orcid":false,"given":"Min","family":"Jiang","sequence":"additional","affiliation":[{"name":"Westlake Laboratory of Life Sciences and Biomedicine","place":["Hangzhou, China"]},{"name":"Institute of Biology, Westlake Institute for Advanced Study","place":["Hangzhou, China"]},{"name":"Key Laboratory of Growth Regulation and Translational Research of Zhejiang Province, School of Life Sciences, Westlake University","place":["Hangzhou, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1999-8451","authenticated-orcid":false,"given":"Zhen","family":"Yan","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Gene Expression, School of Life Sciences, Westlake University","place":["Hangzhou, Zhejiang, China"]},{"name":"Zhejiang Key Laboratory of Structural Biology, School of Life Sciences, Westlake University","place":["Hangzhou, China"]},{"name":"Westlake Laboratory of Life Sciences and Biomedicine","place":["Hangzhou, China"]},{"name":"Institute of Biology, Westlake Institute for Advanced Study","place":["Hangzhou, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9869-7691","authenticated-orcid":false,"given":"Jianping","family":"Wu","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Gene Expression, School of Life Sciences, Westlake University","place":["Hangzhou, Zhejiang, China"]},{"name":"Zhejiang Key Laboratory of Structural Biology, School of Life Sciences, Westlake University","place":["Hangzhou, China"]},{"name":"Westlake Laboratory of Life Sciences and Biomedicine","place":["Hangzhou, China"]},{"name":"Institute of Biology, Westlake Institute for Advanced Study","place":["Hangzhou, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"341","published-online":{"date-parts":[[2026,1,5]]},"reference":[{"key":"e_1_3_4_1_2","doi-asserted-by":"publisher","DOI":"10.1017\/S0967199400002240"},{"key":"e_1_3_4_2_2","doi-asserted-by":"publisher","DOI":"10.1002\/mrd.10017"},{"key":"e_1_3_4_3_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.221454998"},{"key":"e_1_3_4_4_2","doi-asserted-by":"publisher","DOI":"10.1186\/1477-7827-1-53"},{"key":"e_1_3_4_5_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M701083200"},{"key":"e_1_3_4_6_2","doi-asserted-by":"publisher","DOI":"10.1095\/biolreprod.109.077107"},{"key":"e_1_3_4_7_2","doi-asserted-by":"publisher","DOI":"10.1038\/ncomms1153"},{"key":"e_1_3_4_8_2","doi-asserted-by":"publisher","DOI":"10.7554\/eLife.23082"},{"key":"e_1_3_4_9_2","doi-asserted-by":"publisher","DOI":"10.1038\/35098027"},{"key":"e_1_3_4_10_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-021-03742-6"},{"key":"e_1_3_4_11_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-025-65952-0"},{"key":"e_1_3_4_12_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2019.03.047"},{"key":"e_1_3_4_13_2","doi-asserted-by":"publisher","DOI":"10.1186\/s12958-017-0281-2"},{"key":"e_1_3_4_14_2","doi-asserted-by":"publisher","DOI":"10.1152\/physiol.00049.2009"},{"key":"e_1_3_4_15_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cophys.2018.02.004"},{"key":"e_1_3_4_16_2","doi-asserted-by":"publisher","DOI":"10.1152\/physiol.00028.2022"},{"key":"e_1_3_4_17_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.2136654100"},{"key":"e_1_3_4_18_2","doi-asserted-by":"publisher","DOI":"10.1095\/biolreprod.107.060186"},{"key":"e_1_3_4_19_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0610286104"},{"key":"e_1_3_4_20_2","doi-asserted-by":"publisher","DOI":"10.1136\/jmg.2006.045765"},{"key":"e_1_3_4_21_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.ajhg.2009.03.004"},{"key":"e_1_3_4_22_2","doi-asserted-by":"publisher","DOI":"10.1038\/ejhg.2010.108"},{"key":"e_1_3_4_23_2","doi-asserted-by":"publisher","DOI":"10.1093\/humrep\/dey278"},{"key":"e_1_3_4_24_2","doi-asserted-by":"publisher","DOI":"10.1093\/humrep\/dey377"},{"key":"e_1_3_4_25_2","doi-asserted-by":"publisher","DOI":"10.1038\/sj.ejhg.5200991"},{"key":"e_1_3_4_26_2","doi-asserted-by":"publisher","DOI":"10.7150\/thno.51869"},{"key":"e_1_3_4_27_2","doi-asserted-by":"publisher","DOI":"10.1002\/mgg3.1579"},{"key":"e_1_3_4_28_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2014.02.056"},{"key":"e_1_3_4_29_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.celrep.2021.110226"},{"key":"e_1_3_4_30_2","doi-asserted-by":"publisher","DOI":"10.7554\/eLife.62043"},{"key":"e_1_3_4_31_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-022-31050-8"},{"key":"e_1_3_4_32_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.2304409120"},{"key":"e_1_3_4_33_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.2536658100"},{"key":"e_1_3_4_34_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M501430200"},{"key":"e_1_3_4_35_2","doi-asserted-by":"publisher","DOI":"10.1083\/jcb.99.5.1735"},{"key":"e_1_3_4_36_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jmb.2005.04.002"},{"key":"e_1_3_4_37_2","doi-asserted-by":"publisher","DOI":"10.1126\/sciadv.adl1126"},{"key":"e_1_3_4_38_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.aar1968"},{"key":"e_1_3_4_39_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.contraception.2010.03.017"},{"key":"e_1_3_4_40_2","doi-asserted-by":"publisher","DOI":"10.1373\/clinchem.2018.295089"},{"key":"e_1_3_4_41_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0006844"},{"key":"e_1_3_4_42_2","doi-asserted-by":"publisher","DOI":"10.1038\/nature19321"},{"key":"e_1_3_4_43_2","doi-asserted-by":"publisher","DOI":"10.4103\/1008-682X.153303"},{"key":"e_1_3_4_44_2","doi-asserted-by":"crossref","unstructured":"G. M. Dias Thiol-disulfide proteins of stallion epididymal spermatozoa. Anim. Reprod. Sci. 145 29\u201339 (2014) 10.1016\/j.anireprosci.2013.12.007.","DOI":"10.1016\/j.anireprosci.2013.12.007"},{"key":"e_1_3_4_45_2","doi-asserted-by":"crossref","unstructured":"Q. Xu The atomic coordinates of the mouse CatSper dimer. PDB. https:\/\/www.rcsb.org\/structure\/9UBW. Deposited 10 April 2025.","DOI":"10.2210\/pdb9ubw\/pdb"},{"key":"e_1_3_4_46_2","unstructured":"Q. Xu The overall cryo-EM map of the mouse CatSper dimer. EMDB. https:\/\/www.ebi.ac.uk\/emdb\/EMD-64017. Deposited 10 April 2025."},{"key":"e_1_3_4_47_2","unstructured":"Q. Xu The local cryo-EM map of CATSPER\u03b2 of Monomer A of the CatSper dimer. EMDB. https:\/\/www.ebi.ac.uk\/emdb\/EMD-64018. Deposited 10 April 2025."},{"key":"e_1_3_4_48_2","unstructured":"Q. Xu The local cryo-EM map of CATSPER\u03b3 of Monomer A of the CatSper dimer. EMDB. https:\/\/www.ebi.ac.uk\/emdb\/EMD-64019. Deposited 10 April 2025."},{"key":"e_1_3_4_49_2","unstructured":"Q. Xu The local cryo-EM map of CATSPER\u03b4 of Monomer A of the CatSper dimer. EMDB. https:\/\/www.ebi.ac.uk\/emdb\/EMD-64020. Deposited 10 April 2025."},{"key":"e_1_3_4_50_2","unstructured":"Q. Xu The local cryo-EM map of CATSPER\u03b5 of Monomer A of the CatSper dimer. EMDB. https:\/\/www.ebi.ac.uk\/emdb\/EMD-64021. Deposited 10 April 2025."},{"key":"e_1_3_4_51_2","unstructured":"Q. Xu The local cryo-EM map of CATSPER\u03b2 of Monomer B of the CatSper dimer. EMDB. https:\/\/www.ebi.ac.uk\/emdb\/EMD-64022. Deposited 10 April 2025."},{"key":"e_1_3_4_52_2","unstructured":"Q. Xu The local cryo-EM map of CATSPER\u03b3 of Monomer B of the CatSper dimer. EMDB. https:\/\/www.ebi.ac.uk\/emdb\/EMD-64023. Deposited 10 April 2025."},{"key":"e_1_3_4_53_2","unstructured":"Q. Xu The local cryo-EM map of CATSPER\u03b4 of Monomer B of the CatSper dimer. EMDB. https:\/\/www.ebi.ac.uk\/emdb\/EMD-64024. Deposited 10 April 2025."},{"key":"e_1_3_4_54_2","unstructured":"Q. Xu The local cryo-EM map of CATSPER\u03b5 of Monomer B of the CatSper dimer. EMDB. https:\/\/www.ebi.ac.uk\/emdb\/EMD-64025. Deposited 10 April 2025."},{"key":"e_1_3_4_55_2","unstructured":"Q. Xu The local cryo-EM map of the transmembrane region of the CatSper dimer. EMDB. https:\/\/www.ebi.ac.uk\/emdb\/EMD-64026. Deposited 10 April 2025."},{"key":"e_1_3_4_56_2","unstructured":"Q. Xu The local cryo-EM map of the \u03b7-\u03b7 interface of the CatSper dimer. EMDB. https:\/\/www.ebi.ac.uk\/emdb\/EMD-64027. Deposited 10 April 2025."},{"key":"e_1_3_4_57_2","doi-asserted-by":"crossref","unstructured":"Q. Xu The atomic coordinates of the mouse CatSper trimer. PDB. https:\/\/www.rcsb.org\/structure\/9UBX. Deposited 10 April 2025.","DOI":"10.2210\/pdb9ubx\/pdb"},{"key":"e_1_3_4_58_2","unstructured":"Q. Xu The overall cryo-EM map of the mouse CatSper trimer. EMDB. https:\/\/www.ebi.ac.uk\/emdb\/EMD-64028. Deposited 10 April 2025."},{"key":"e_1_3_4_59_2","unstructured":"Q. Xu The local cryo-EM map of the extracellular region of Monomer A of the CatSper trimer. EMDB. https:\/\/www.ebi.ac.uk\/emdb\/EMD-64029. Deposited 10 April 2025."},{"key":"e_1_3_4_60_2","unstructured":"Q. Xu The local cryo-EM map of the extracellular region of Monomers B and A' of the CatSper trimer. EMDB. https:\/\/www.ebi.ac.uk\/emdb\/EMD-64030. Deposited 10 April 2025."},{"key":"e_1_3_4_61_2","unstructured":"Q. Xu The local cryo-EM map of the \u03b3-\u03b3 interface of the CatSper trimer. EMDB. https:\/\/www.ebi.ac.uk\/emdb\/EMD-64031. Deposited 10 April 2025."},{"key":"e_1_3_4_62_2","unstructured":"Q. Xu Proteomic analysis of the sperm from Catsperh\/Tmem262 knockout mice. iProX. https:\/\/www.iprox.cn\/page\/project.html?id=IPX0014152000. Deposited 6 November 2025."},{"key":"e_1_3_4_63_2","unstructured":"Q. Zhao Proteomic analysis of the sperm from WT and Armh2 knockout mice. iProX. https:\/\/www.iprox.cn\/\/page\/project.html?id=IPX0013365000. Deposited 9 September 2025."},{"key":"e_1_3_4_64_2","unstructured":"S. Lin The atomic coordinates of the mouse CatSper monomer. PDB. https:\/\/www.rcsb.org\/structure\/7EEB. Deposited 18 March 2021."},{"key":"e_1_3_4_65_2","unstructured":"Y. Zhao Sub-tomographic average of CatSper array from mouse wild type sperm flagella. EMDB. https:\/\/www.ebi.ac.uk\/emdb\/EMD-24210. Deposited 8 June 2022."}],"container-title":["Proceedings of the National Academy of Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pnas.org\/doi\/pdf\/10.1073\/pnas.2510754123","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,1,5]],"date-time":"2026-01-05T20:02:56Z","timestamp":1767643376000},"score":1,"resource":{"primary":{"URL":"https:\/\/pnas.org\/doi\/10.1073\/pnas.2510754123"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,1,5]]},"references-count":65,"journal-issue":{"issue":"2","published-print":{"date-parts":[[2026,1,13]]}},"alternative-id":["10.1073\/pnas.2510754123"],"URL":"https:\/\/doi.org\/10.1073\/pnas.2510754123","relation":{},"ISSN":["0027-8424","1091-6490"],"issn-type":[{"value":"0027-8424","type":"print"},{"value":"1091-6490","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,1,5]]},"assertion":[{"value":"2025-05-03","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-11-25","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-01-05","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e2510754123"}}