[{"scopus_import":"1","keyword":["Molecular Biology","Structural Biology"],"article_processing_charge":"Yes (via OA deal)","has_accepted_license":"1","day":"01","citation":{"chicago":"Kampjut, Domen, and Leonid A Sazanov. “Structure of Respiratory Complex I – An Emerging Blueprint for the Mechanism.” Current Opinion in Structural Biology. Elsevier, 2022. https://doi.org/10.1016/j.sbi.2022.102350.","mla":"Kampjut, Domen, and Leonid A. Sazanov. “Structure of Respiratory Complex I – An Emerging Blueprint for the Mechanism.” Current Opinion in Structural Biology, vol. 74, 102350, Elsevier, 2022, doi:10.1016/j.sbi.2022.102350.","short":"D. Kampjut, L.A. Sazanov, Current Opinion in Structural Biology 74 (2022).","ista":"Kampjut D, Sazanov LA. 2022. Structure of respiratory complex I – An emerging blueprint for the mechanism. Current Opinion in Structural Biology. 74, 102350.","apa":"Kampjut, D., & Sazanov, L. A. (2022). Structure of respiratory complex I – An emerging blueprint for the mechanism. Current Opinion in Structural Biology. Elsevier. https://doi.org/10.1016/j.sbi.2022.102350","ieee":"D. Kampjut and L. A. Sazanov, “Structure of respiratory complex I – An emerging blueprint for the mechanism,” Current Opinion in Structural Biology, vol. 74. Elsevier, 2022.","ama":"Kampjut D, Sazanov LA. Structure of respiratory complex I – An emerging blueprint for the mechanism. Current Opinion in Structural Biology. 2022;74. doi:10.1016/j.sbi.2022.102350"},"publication":"Current Opinion in Structural Biology","article_type":"original","date_published":"2022-06-01T00:00:00Z","type":"journal_article","abstract":[{"lang":"eng","text":"Complex I is one of the major respiratory complexes, conserved from bacteria to mammals. It oxidises NADH, reduces quinone and pumps protons across the membrane, thus playing a central role in the oxidative energy metabolism. In this review we discuss our current state of understanding the structure of complex I from various species of mammals, plants, fungi, and bacteria, as well as of several complex I-related proteins. By comparing the structural evidence from these systems in different redox states and data from mutagenesis and molecular simulations, we formulate the mechanisms of electron transfer and proton pumping and explain how they are conformationally and electrostatically coupled. Finally, we discuss the structural basis of the deactivation phenomenon in mammalian complex I."}],"user_id":"4359f0d1-fa6c-11eb-b949-802e58b17ae8","_id":"11167","intvolume":" 74","status":"public","title":"Structure of respiratory complex I – An emerging blueprint for the mechanism","ddc":["570"],"oa_version":"Published Version","file":[{"date_updated":"2022-08-05T05:56:03Z","date_created":"2022-08-05T05:56:03Z","success":1,"checksum":"72bdde48853643a32d42b75f54965c44","file_id":"11725","relation":"main_file","creator":"dernst","content_type":"application/pdf","file_size":815607,"file_name":"2022_CurrentOpStructBiology_Kampjut.pdf","access_level":"open_access"}],"publication_identifier":{"issn":["0959-440X"]},"month":"06","external_id":{"isi":["000829029500020"],"pmid":["35316665"]},"tmp":{"name":"Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)","legal_code_url":"https://creativecommons.org/licenses/by/4.0/legalcode","short":"CC BY (4.0)","image":"/images/cc_by.png"},"oa":1,"quality_controlled":"1","isi":1,"doi":"10.1016/j.sbi.2022.102350","language":[{"iso":"eng"}],"article_number":"102350","file_date_updated":"2022-08-05T05:56:03Z","pmid":1,"year":"2022","department":[{"_id":"LeSa"}],"publisher":"Elsevier","publication_status":"published","author":[{"full_name":"Kampjut, Domen","first_name":"Domen","last_name":"Kampjut","id":"37233050-F248-11E8-B48F-1D18A9856A87"},{"last_name":"Sazanov","first_name":"Leonid A","orcid":"0000-0002-0977-7989","id":"338D39FE-F248-11E8-B48F-1D18A9856A87","full_name":"Sazanov, Leonid A"}],"volume":74,"date_updated":"2023-08-03T06:31:06Z","date_created":"2022-04-15T09:32:35Z"},{"pmid":1,"year":"2022","acknowledgement":"We thank Dr, Luke Formosa (Department of Biochemistry and Molecular Biology, Monash Biomedicine Discovery Institute, Monash University, Melbourne, Australia) for his valuable advice and assistance on NDUFA10 molecular studies and Dr. Francesc Canals and his team (Proteomics Laboratory, Vall d’Hebron Institute of Oncology [VHIO], Universitat Autònoma de Barcelona, Barcelona, Spain) for their assistance with LC-MS/MS analyses. This work was supported by the Spanish Ministry of Industry, Economy and Competitiveness [grants BFU2014-52618-R, SAF2017-87506, and PID2020-112929RB-I00 to Y.C.], by the Spanish Instituto de Salud Carlos III [grants PI21/00554 and PMP15/00025 to R.M.], co-financed by the European Regional Development Fund (ERDF), and by an NHMRC Project grant to M.R. (GNT1164459).\r\n","department":[{"_id":"LeSa"}],"publisher":"Springer Nature","publication_status":"published","author":[{"full_name":"Molina-Granada, David","last_name":"Molina-Granada","first_name":"David"},{"full_name":"González-Vioque, Emiliano","last_name":"González-Vioque","first_name":"Emiliano"},{"first_name":"Marris G.","last_name":"Dibley","full_name":"Dibley, Marris G."},{"first_name":"Raquel","last_name":"Cabrera-Pérez","full_name":"Cabrera-Pérez, Raquel"},{"last_name":"Vallbona-Garcia","first_name":"Antoni","full_name":"Vallbona-Garcia, Antoni"},{"full_name":"Torres-Torronteras, Javier","first_name":"Javier","last_name":"Torres-Torronteras"},{"id":"338D39FE-F248-11E8-B48F-1D18A9856A87","orcid":"0000-0002-0977-7989","first_name":"Leonid A","last_name":"Sazanov","full_name":"Sazanov, Leonid A"},{"full_name":"Ryan, Michael T.","first_name":"Michael T.","last_name":"Ryan"},{"last_name":"Cámara","first_name":"Yolanda","full_name":"Cámara, Yolanda"},{"last_name":"Martí","first_name":"Ramon","full_name":"Martí, Ramon"}],"volume":5,"date_created":"2022-07-10T22:01:52Z","date_updated":"2023-08-03T11:51:58Z","article_number":"620","file_date_updated":"2022-07-13T07:44:58Z","tmp":{"name":"Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)","legal_code_url":"https://creativecommons.org/licenses/by/4.0/legalcode","short":"CC BY (4.0)","image":"/images/cc_by.png"},"oa":1,"external_id":{"isi":["000815098500002"],"pmid":[" 35739187"]},"isi":1,"quality_controlled":"1","doi":"10.1038/s42003-022-03568-6","language":[{"iso":"eng"}],"publication_identifier":{"eissn":["23993642"]},"month":"06","user_id":"4359f0d1-fa6c-11eb-b949-802e58b17ae8","_id":"11551","intvolume":" 5","title":"Most mitochondrial dGTP is tightly bound to respiratory complex I through the NDUFA10 subunit","status":"public","ddc":["570"],"oa_version":"Published Version","file":[{"date_created":"2022-07-13T07:44:58Z","date_updated":"2022-07-13T07:44:58Z","checksum":"965f88bbcef3fd0c3e121340555c4467","success":1,"relation":"main_file","file_id":"11571","file_size":2335369,"content_type":"application/pdf","creator":"kschuh","file_name":"2022_communicationsbiology_Molina-Granada.pdf","access_level":"open_access"}],"type":"journal_article","issue":"1","abstract":[{"text":"Imbalanced mitochondrial dNTP pools are known players in the pathogenesis of multiple human diseases. Here we show that, even under physiological conditions, dGTP is largely overrepresented among other dNTPs in mitochondria of mouse tissues and human cultured cells. In addition, a vast majority of mitochondrial dGTP is tightly bound to NDUFA10, an accessory subunit of complex I of the mitochondrial respiratory chain. NDUFA10 shares a deoxyribonucleoside kinase (dNK) domain with deoxyribonucleoside kinases in the nucleotide salvage pathway, though no specific function beyond stabilizing the complex I holoenzyme has been described for this subunit. We mutated the dNK domain of NDUFA10 in human HEK-293T cells while preserving complex I assembly and activity. The NDUFA10E160A/R161A shows reduced dGTP binding capacity in vitro and leads to a 50% reduction in mitochondrial dGTP content, proving that most dGTP is directly bound to the dNK domain of NDUFA10. This interaction may represent a hitherto unknown mechanism regulating mitochondrial dNTP availability and linking oxidative metabolism to DNA maintenance.","lang":"eng"}],"citation":{"ieee":"D. Molina-Granada et al., “Most mitochondrial dGTP is tightly bound to respiratory complex I through the NDUFA10 subunit,” Communications Biology, vol. 5, no. 1. Springer Nature, 2022.","apa":"Molina-Granada, D., González-Vioque, E., Dibley, M. G., Cabrera-Pérez, R., Vallbona-Garcia, A., Torres-Torronteras, J., … Martí, R. (2022). Most mitochondrial dGTP is tightly bound to respiratory complex I through the NDUFA10 subunit. Communications Biology. Springer Nature. https://doi.org/10.1038/s42003-022-03568-6","ista":"Molina-Granada D, González-Vioque E, Dibley MG, Cabrera-Pérez R, Vallbona-Garcia A, Torres-Torronteras J, Sazanov LA, Ryan MT, Cámara Y, Martí R. 2022. Most mitochondrial dGTP is tightly bound to respiratory complex I through the NDUFA10 subunit. Communications Biology. 5(1), 620.","ama":"Molina-Granada D, González-Vioque E, Dibley MG, et al. Most mitochondrial dGTP is tightly bound to respiratory complex I through the NDUFA10 subunit. Communications Biology. 2022;5(1). doi:10.1038/s42003-022-03568-6","chicago":"Molina-Granada, David, Emiliano González-Vioque, Marris G. Dibley, Raquel Cabrera-Pérez, Antoni Vallbona-Garcia, Javier Torres-Torronteras, Leonid A Sazanov, Michael T. Ryan, Yolanda Cámara, and Ramon Martí. “Most Mitochondrial DGTP Is Tightly Bound to Respiratory Complex I through the NDUFA10 Subunit.” Communications Biology. Springer Nature, 2022. https://doi.org/10.1038/s42003-022-03568-6.","short":"D. Molina-Granada, E. González-Vioque, M.G. Dibley, R. Cabrera-Pérez, A. Vallbona-Garcia, J. Torres-Torronteras, L.A. Sazanov, M.T. Ryan, Y. Cámara, R. Martí, Communications Biology 5 (2022).","mla":"Molina-Granada, David, et al. “Most Mitochondrial DGTP Is Tightly Bound to Respiratory Complex I through the NDUFA10 Subunit.” Communications Biology, vol. 5, no. 1, 620, Springer Nature, 2022, doi:10.1038/s42003-022-03568-6."},"publication":"Communications Biology","date_published":"2022-06-23T00:00:00Z","scopus_import":"1","article_processing_charge":"No","has_accepted_license":"1","day":"23"},{"language":[{"iso":"eng"}],"doi":"10.1093/jmicro/dfac037","isi":1,"quality_controlled":"1","oa":1,"tmp":{"name":"Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)","legal_code_url":"https://creativecommons.org/licenses/by/4.0/legalcode","short":"CC BY (4.0)","image":"/images/cc_by.png"},"external_id":{"pmid":["35861182"],"isi":["000837950900001"]},"month":"10","publication_identifier":{"eissn":["2050-5701"],"issn":["2050-5698"]},"date_created":"2022-07-25T10:04:58Z","date_updated":"2023-08-03T12:13:37Z","volume":71,"author":[{"full_name":"Gerle, Christoph","last_name":"Gerle","first_name":"Christoph"},{"full_name":"Kishikawa, Jun-ichi","first_name":"Jun-ichi","last_name":"Kishikawa"},{"last_name":"Yamaguchi","first_name":"Tomoko","full_name":"Yamaguchi, Tomoko"},{"first_name":"Atsuko","last_name":"Nakanishi","full_name":"Nakanishi, Atsuko"},{"first_name":"Mehmet Orkun","last_name":"Çoruh","id":"d25163e5-8d53-11eb-a251-e6dd8ea1b8ef","orcid":"0000-0002-3219-2022","full_name":"Çoruh, Mehmet Orkun"},{"first_name":"Fumiaki","last_name":"Makino","full_name":"Makino, Fumiaki"},{"first_name":"Tomoko","last_name":"Miyata","full_name":"Miyata, Tomoko"},{"last_name":"Kawamoto","first_name":"Akihiro","full_name":"Kawamoto, Akihiro"},{"full_name":"Yokoyama, Ken","first_name":"Ken","last_name":"Yokoyama"},{"full_name":"Namba, Keiichi","first_name":"Keiichi","last_name":"Namba"},{"full_name":"Kurisu, Genji","first_name":"Genji","last_name":"Kurisu"},{"full_name":"Kato, Takayuki","last_name":"Kato","first_name":"Takayuki"}],"publication_status":"published","department":[{"_id":"LeSa"}],"publisher":"Oxford University Press","year":"2022","acknowledgement":"Cyclic Innovation for Clinical Empowerment (JP17pc0101020 from Japan Agency for Medical Research and Development (AMED) to K.N. and G.K.); Platform Project for Supporting Drug Discovery and Life Science Research (Basis for Supporting Innovative Drug Discovery and Life Science Research) from AMED (JP20am0101117 to K.N., JP16K07266 to Atsunori Oshima and C.G., JP22ama121001j0001 to Masaki Yamamoto, G.K., T.K. and C.G.); a JSPS KAHKENHI\r\ngrant (20K06514 to J.K.) and a Grant-in-aid for JSPS fellows (20J00162 to A.N.).\r\nWe are grateful for initiation and scientific support from Matthias Rogner, Marc M. Nowaczyk, Anna Frank and ̈Yuko Misumi for the PSI monomer project and also would like to thank Hideki Shigematsu for critical reading of the manuscript. And we are indebted to the two anonymous reviewers who helped us to improve our manuscript.","pmid":1,"file_date_updated":"2023-02-03T08:34:48Z","date_published":"2022-10-01T00:00:00Z","article_type":"original","page":"249-261","publication":"Microscopy","citation":{"ama":"Gerle C, Kishikawa J, Yamaguchi T, et al. Structures of multisubunit membrane complexes with the CRYO ARM 200. Microscopy. 2022;71(5):249-261. doi:10.1093/jmicro/dfac037","ieee":"C. Gerle et al., “Structures of multisubunit membrane complexes with the CRYO ARM 200,” Microscopy, vol. 71, no. 5. Oxford University Press, pp. 249–261, 2022.","apa":"Gerle, C., Kishikawa, J., Yamaguchi, T., Nakanishi, A., Çoruh, M. O., Makino, F., … Kato, T. (2022). Structures of multisubunit membrane complexes with the CRYO ARM 200. Microscopy. Oxford University Press. https://doi.org/10.1093/jmicro/dfac037","ista":"Gerle C, Kishikawa J, Yamaguchi T, Nakanishi A, Çoruh MO, Makino F, Miyata T, Kawamoto A, Yokoyama K, Namba K, Kurisu G, Kato T. 2022. Structures of multisubunit membrane complexes with the CRYO ARM 200. Microscopy. 71(5), 249–261.","short":"C. Gerle, J. Kishikawa, T. Yamaguchi, A. Nakanishi, M.O. Çoruh, F. Makino, T. Miyata, A. Kawamoto, K. Yokoyama, K. Namba, G. Kurisu, T. Kato, Microscopy 71 (2022) 249–261.","mla":"Gerle, Christoph, et al. “Structures of Multisubunit Membrane Complexes with the CRYO ARM 200.” Microscopy, vol. 71, no. 5, Oxford University Press, 2022, pp. 249–61, doi:10.1093/jmicro/dfac037.","chicago":"Gerle, Christoph, Jun-ichi Kishikawa, Tomoko Yamaguchi, Atsuko Nakanishi, Mehmet Orkun Çoruh, Fumiaki Makino, Tomoko Miyata, et al. “Structures of Multisubunit Membrane Complexes with the CRYO ARM 200.” Microscopy. Oxford University Press, 2022. https://doi.org/10.1093/jmicro/dfac037."},"day":"01","article_processing_charge":"No","has_accepted_license":"1","keyword":["Radiology","Nuclear Medicine and imaging","Instrumentation","Structural Biology"],"scopus_import":"1","file":[{"file_id":"12498","relation":"main_file","date_created":"2023-02-03T08:34:48Z","date_updated":"2023-02-03T08:34:48Z","success":1,"checksum":"23b51c163636bf9313f7f0818312e67e","file_name":"2022_Microscopy_Gerle.pdf","access_level":"open_access","creator":"dernst","file_size":7812696,"content_type":"application/pdf"}],"oa_version":"Published Version","ddc":["570"],"title":"Structures of multisubunit membrane complexes with the CRYO ARM 200","status":"public","intvolume":" 71","user_id":"4359f0d1-fa6c-11eb-b949-802e58b17ae8","_id":"11648","abstract":[{"text":"Progress in structural membrane biology has been significantly accelerated by the ongoing 'Resolution Revolution' in cryo electron microscopy (cryo-EM). In particular, structure determination by single particle analysis has evolved into the most powerful method for atomic model building of multisubunit membrane protein complexes. This has created an ever increasing demand in cryo-EM machine time, which to satisfy is in need of new and affordable cryo electron microscopes. Here, we review our experience in using the JEOL CRYO ARM 200 prototype for the structure determination by single particle analysis of three different multisubunit membrane complexes: the Thermus thermophilus V-type ATPase VO complex, the Thermosynechococcus elongatus photosystem I monomer and the flagellar motor LP-ring from Salmonella enterica.","lang":"eng"}],"issue":"5","type":"journal_article"},{"scopus_import":"1","keyword":["Multidisciplinary"],"has_accepted_license":"1","article_processing_charge":"No","day":"22","citation":{"ama":"Kravchuk V, Petrova O, Kampjut D, Wojciechowska-Bason A, Breese Z, Sazanov LA. A universal coupling mechanism of respiratory complex I. Nature. 2022;609(7928):808-814. doi:10.1038/s41586-022-05199-7","ieee":"V. Kravchuk, O. Petrova, D. Kampjut, A. Wojciechowska-Bason, Z. Breese, and L. A. Sazanov, “A universal coupling mechanism of respiratory complex I,” Nature, vol. 609, no. 7928. Springer Nature, pp. 808–814, 2022.","apa":"Kravchuk, V., Petrova, O., Kampjut, D., Wojciechowska-Bason, A., Breese, Z., & Sazanov, L. A. (2022). A universal coupling mechanism of respiratory complex I. Nature. Springer Nature. https://doi.org/10.1038/s41586-022-05199-7","ista":"Kravchuk V, Petrova O, Kampjut D, Wojciechowska-Bason A, Breese Z, Sazanov LA. 2022. A universal coupling mechanism of respiratory complex I. Nature. 609(7928), 808–814.","short":"V. Kravchuk, O. Petrova, D. Kampjut, A. Wojciechowska-Bason, Z. Breese, L.A. Sazanov, Nature 609 (2022) 808–814.","mla":"Kravchuk, Vladyslav, et al. “A Universal Coupling Mechanism of Respiratory Complex I.” Nature, vol. 609, no. 7928, Springer Nature, 2022, pp. 808–14, doi:10.1038/s41586-022-05199-7.","chicago":"Kravchuk, Vladyslav, Olga Petrova, Domen Kampjut, Anna Wojciechowska-Bason, Zara Breese, and Leonid A Sazanov. “A Universal Coupling Mechanism of Respiratory Complex I.” Nature. Springer Nature, 2022. https://doi.org/10.1038/s41586-022-05199-7."},"publication":"Nature","page":"808-814","article_type":"original","date_published":"2022-09-22T00:00:00Z","type":"journal_article","issue":"7928","abstract":[{"text":"Complex I is the first enzyme in the respiratory chain, which is responsible for energy production in mitochondria and bacteria1. Complex I couples the transfer of two electrons from NADH to quinone and the translocation of four protons across the membrane2, but the coupling mechanism remains contentious. Here we present cryo-electron microscopy structures of Escherichia coli complex I (EcCI) in different redox states, including catalytic turnover. EcCI exists mostly in the open state, in which the quinone cavity is exposed to the cytosol, allowing access for water molecules, which enable quinone movements. Unlike the mammalian paralogues3, EcCI can convert to the closed state only during turnover, showing that closed and open states are genuine turnover intermediates. The open-to-closed transition results in the tightly engulfed quinone cavity being connected to the central axis of the membrane arm, a source of substrate protons. Consistently, the proportion of the closed state increases with increasing pH. We propose a detailed but straightforward and robust mechanism comprising a ‘domino effect’ series of proton transfers and electrostatic interactions: the forward wave (‘dominoes stacking’) primes the pump, and the reverse wave (‘dominoes falling’) results in the ejection of all pumped protons from the distal subunit NuoL. This mechanism explains why protons exit exclusively from the NuoL subunit and is supported by our mutagenesis data. We contend that this is a universal coupling mechanism of complex I and related enzymes.","lang":"eng"}],"user_id":"4359f0d1-fa6c-11eb-b949-802e58b17ae8","_id":"12138","intvolume":" 609","ddc":["572"],"title":"A universal coupling mechanism of respiratory complex I","status":"public","file":[{"file_id":"13104","relation":"main_file","date_created":"2023-05-30T17:05:31Z","date_updated":"2023-05-30T17:05:31Z","success":1,"checksum":"d42a93e24f59e883ef0b5429832391d0","file_name":"EcCxI_manuscript_rev3_noSI_updated_withFigs_opt.pdf","access_level":"open_access","creator":"lsazanov","content_type":"application/pdf","file_size":1425655},{"relation":"main_file","file_id":"13105","checksum":"5422bc0a73b3daadafa262c7ea6deae3","success":1,"date_created":"2023-05-30T17:07:05Z","date_updated":"2023-05-30T17:07:05Z","access_level":"open_access","file_name":"EcCxI_manuscript_rev3_SI_All_opt_upd.pdf","content_type":"application/pdf","file_size":9842513,"creator":"lsazanov"}],"oa_version":"Submitted Version","publication_identifier":{"issn":["0028-0836"],"eissn":["1476-4687"]},"month":"09","external_id":{"isi":["000854788200001"],"pmid":["36104567"]},"oa":1,"project":[{"name":"Structural characterization of E. coli complex I: an important mechanistic model","grant_number":"25541","_id":"238A0A5A-32DE-11EA-91FC-C7463DDC885E"},{"grant_number":"101020697","_id":"627abdeb-2b32-11ec-9570-ec31a97243d3","name":"Structure and mechanism of respiratory chain molecular machines","call_identifier":"H2020"}],"isi":1,"quality_controlled":"1","doi":"10.1038/s41586-022-05199-7","language":[{"iso":"eng"}],"acknowledged_ssus":[{"_id":"EM-Fac"},{"_id":"LifeSc"},{"_id":"ScienComp"}],"ec_funded":1,"file_date_updated":"2023-05-30T17:07:05Z","pmid":1,"year":"2022","acknowledgement":"This research was supported by the Scientific Service Units (SSU) of IST Austria through resources provided by the Electron Microscopy Facility (EMF), the Life Science Facility (LSF) and the IST high-performance computing cluster. We thank V.-V. Hodirnau from IST Austria EMF, M. Babiak from CEITEC for assistance with collecting cryo-EM data and A. Charnagalov for the assistance with protein purification. V.K. was a recipient of a DOC Fellowship of the Austrian Academy of Sciences at the Institute of Science and Technology, Austria. V.K. and O.P. are funded by the ERC Advanced Grant 101020697 RESPICHAIN to L.S. This work was also supported by the Medical Research Council (UK).","publisher":"Springer Nature","department":[{"_id":"LeSa"}],"publication_status":"published","related_material":{"link":[{"url":"https://doi.org/10.1038/s41586-022-05457-8","relation":"erratum"},{"description":"News on ISTA website","relation":"press_release","url":"https://ista.ac.at/en/news/proton-dominos-kick-off-life/"}],"record":[{"relation":"dissertation_contains","status":"public","id":"12781"}]},"author":[{"full_name":"Kravchuk, Vladyslav","id":"4D62F2A6-F248-11E8-B48F-1D18A9856A87","first_name":"Vladyslav","last_name":"Kravchuk"},{"id":"5D8C9660-5D49-11EA-8188-567B3DDC885E","last_name":"Petrova","first_name":"Olga","full_name":"Petrova, Olga"},{"last_name":"Kampjut","first_name":"Domen","id":"37233050-F248-11E8-B48F-1D18A9856A87","full_name":"Kampjut, Domen"},{"full_name":"Wojciechowska-Bason, Anna","last_name":"Wojciechowska-Bason","first_name":"Anna"},{"full_name":"Breese, Zara","first_name":"Zara","last_name":"Breese"},{"full_name":"Sazanov, Leonid A","last_name":"Sazanov","first_name":"Leonid A","orcid":"0000-0002-0977-7989","id":"338D39FE-F248-11E8-B48F-1D18A9856A87"}],"volume":609,"date_created":"2023-01-12T12:04:33Z","date_updated":"2023-08-04T08:54:52Z"},{"language":[{"iso":"eng"}],"doi":"10.3389/fimmu.2022.965446","isi":1,"quality_controlled":"1","oa":1,"tmp":{"name":"Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)","legal_code_url":"https://creativecommons.org/licenses/by/4.0/legalcode","short":"CC BY (4.0)","image":"/images/cc_by.png"},"external_id":{"isi":["000862479100001"]},"publication_identifier":{"issn":["1664-3224"]},"month":"09","volume":13,"date_updated":"2023-08-04T09:49:24Z","date_created":"2023-01-16T09:56:57Z","author":[{"first_name":"Dmitri","last_name":"Dormeshkin","full_name":"Dormeshkin, Dmitri"},{"last_name":"Shapira","first_name":"Michail","full_name":"Shapira, Michail"},{"full_name":"Dubovik, Simon","first_name":"Simon","last_name":"Dubovik"},{"last_name":"Kavaleuski","first_name":"Anton","orcid":"0000-0003-2091-526X","id":"4968f7ad-eb97-11eb-a6c2-8ed382e8912c","full_name":"Kavaleuski, Anton"},{"first_name":"Mikalai","last_name":"Katsin","full_name":"Katsin, Mikalai"},{"last_name":"Migas","first_name":"Alexandr","full_name":"Migas, Alexandr"},{"first_name":"Alexander","last_name":"Meleshko","full_name":"Meleshko, Alexander"},{"full_name":"Semyonov, Sergei","last_name":"Semyonov","first_name":"Sergei"}],"department":[{"_id":"LeSa"}],"publisher":"Frontiers Media","publication_status":"published","year":"2022","acknowledgement":"The authors declare that this study received funding from Immunofusion. The funder was not involved in the study design, collection, analysis, interpretation of data, the writing of this article or the decision to submit it for publication.","file_date_updated":"2023-01-30T09:22:26Z","article_number":"965446","date_published":"2022-09-16T00:00:00Z","article_type":"original","citation":{"ieee":"D. Dormeshkin et al., “Isolation of an escape-resistant SARS-CoV-2 neutralizing nanobody from a novel synthetic nanobody library,” Frontiers in Immunology, vol. 13. Frontiers Media, 2022.","apa":"Dormeshkin, D., Shapira, M., Dubovik, S., Kavaleuski, A., Katsin, M., Migas, A., … Semyonov, S. (2022). Isolation of an escape-resistant SARS-CoV-2 neutralizing nanobody from a novel synthetic nanobody library. Frontiers in Immunology. Frontiers Media. https://doi.org/10.3389/fimmu.2022.965446","ista":"Dormeshkin D, Shapira M, Dubovik S, Kavaleuski A, Katsin M, Migas A, Meleshko A, Semyonov S. 2022. Isolation of an escape-resistant SARS-CoV-2 neutralizing nanobody from a novel synthetic nanobody library. Frontiers in Immunology. 13, 965446.","ama":"Dormeshkin D, Shapira M, Dubovik S, et al. Isolation of an escape-resistant SARS-CoV-2 neutralizing nanobody from a novel synthetic nanobody library. Frontiers in Immunology. 2022;13. doi:10.3389/fimmu.2022.965446","chicago":"Dormeshkin, Dmitri, Michail Shapira, Simon Dubovik, Anton Kavaleuski, Mikalai Katsin, Alexandr Migas, Alexander Meleshko, and Sergei Semyonov. “Isolation of an Escape-Resistant SARS-CoV-2 Neutralizing Nanobody from a Novel Synthetic Nanobody Library.” Frontiers in Immunology. Frontiers Media, 2022. https://doi.org/10.3389/fimmu.2022.965446.","short":"D. Dormeshkin, M. Shapira, S. Dubovik, A. Kavaleuski, M. Katsin, A. Migas, A. Meleshko, S. Semyonov, Frontiers in Immunology 13 (2022).","mla":"Dormeshkin, Dmitri, et al. “Isolation of an Escape-Resistant SARS-CoV-2 Neutralizing Nanobody from a Novel Synthetic Nanobody Library.” Frontiers in Immunology, vol. 13, 965446, Frontiers Media, 2022, doi:10.3389/fimmu.2022.965446."},"publication":"Frontiers in Immunology","has_accepted_license":"1","article_processing_charge":"No","day":"16","keyword":["Immunology","Immunology and Allergy","COVID-19","SARS-CoV-2","synthetic library","RBD","neutralization nanobody","VHH"],"scopus_import":"1","oa_version":"Published Version","file":[{"relation":"main_file","file_id":"12443","date_created":"2023-01-30T09:22:26Z","date_updated":"2023-01-30T09:22:26Z","checksum":"f8f5d8110710033d0532e7e08bf9dad4","success":1,"file_name":"2022_FrontiersImmunology_Dormeshkin.pdf","access_level":"open_access","file_size":5695892,"content_type":"application/pdf","creator":"dernst"}],"intvolume":" 13","title":"Isolation of an escape-resistant SARS-CoV-2 neutralizing nanobody from a novel synthetic nanobody library","status":"public","ddc":["570"],"user_id":"4359f0d1-fa6c-11eb-b949-802e58b17ae8","_id":"12252","abstract":[{"text":"The COVID−19 pandemic not only resulted in a global crisis, but also accelerated vaccine development and antibody discovery. Herein we report a synthetic humanized VHH library development pipeline for nanomolar-range affinity VHH binders to SARS-CoV-2 variants of concern (VoC) receptor binding domains (RBD) isolation. Trinucleotide-based randomization of CDRs by Kunkel mutagenesis with the subsequent rolling-cycle amplification resulted in more than 1011 diverse phage display library in a manageable for a single person number of electroporation reactions. We identified a number of nanomolar-range affinity VHH binders to SARS-CoV-2 variants of concern (VoC) receptor binding domains (RBD) by screening a novel synthetic humanized antibody library. In order to explore the most robust and fast method for affinity improvement, we performed affinity maturation by CDR1 and CDR2 shuffling and avidity engineering by multivalent trimeric VHH fusion protein construction. As a result, H7-Fc and G12x3-Fc binders were developed with the affinities in nM and pM range respectively. Importantly, these affinities are weakly influenced by most of SARS-CoV-2 VoC mutations and they retain moderate binding to BA.4\\5. The plaque reduction neutralization test (PRNT) resulted in IC50 = 100 ng\\ml and 9.6 ng\\ml for H7-Fc and G12x3-Fc antibodies, respectively, for the emerging Omicron BA.1 variant. Therefore, these VHH could expand the present landscape of SARS-CoV-2 neutralization binders with the therapeutic potential for present and future SARS-CoV-2 variants.","lang":"eng"}],"type":"journal_article"}]