[{"type":"journal_article","abstract":[{"text":"Chemical labeling of proteins with synthetic molecular probes offers the possibility to probe the functions of proteins of interest in living cells. However, the methods for covalently labeling targeted proteins using complementary peptide tag-probe pairs are still limited, irrespective of the versatility of such pairs in biological research. Herein, we report the new CysHis tag-Ni(II) probe pair for the specific covalent labeling of proteins. A broad-range evaluation of the reactivity profiles of the probe and the CysHis peptide tag afforded a tag-probe pair with an optimized and high labeling selectivity and reactivity. In particular, the labeling specificity of this pair was notably improved compared to the previously reported one. This pair was successfully utilized for the fluorescence imaging of membrane proteins on the surfaces of living cells, demonstrating its potential utility in biological research.","lang":"eng"}],"issue":"5","title":"Optimized reaction pair of the CysHis tag and Ni(II)-NTA probe for highly selective chemical labeling of membrane proteins","ddc":["570"],"status":"public","intvolume":" 92","_id":"6659","user_id":"2DF688A6-F248-11E8-B48F-1D18A9856A87","file":[{"file_name":"2019_BCSJ_Zenmyo.pdf","access_level":"open_access","creator":"dernst","file_size":2464903,"content_type":"application/pdf","file_id":"8594","relation":"main_file","date_updated":"2020-10-02T08:49:58Z","date_created":"2020-10-02T08:49:58Z","success":1,"checksum":"186de511d6e0ca93f5d981e2443eb8cd"}],"oa_version":"Published Version","scopus_import":"1","day":"15","article_processing_charge":"No","has_accepted_license":"1","article_type":"original","page":"995-1000","publication":"Bulletin of the Chemical Society of Japan","citation":{"ama":"Zenmyo N, Tokumaru H, Uchinomiya S, et al. Optimized reaction pair of the CysHis tag and Ni(II)-NTA probe for highly selective chemical labeling of membrane proteins. Bulletin of the Chemical Society of Japan. 2019;92(5):995-1000. doi:10.1246/bcsj.20190034","apa":"Zenmyo, N., Tokumaru, H., Uchinomiya, S., Fuchida, H., Tabata, S., Hamachi, I., … Ojida, A. (2019). Optimized reaction pair of the CysHis tag and Ni(II)-NTA probe for highly selective chemical labeling of membrane proteins. Bulletin of the Chemical Society of Japan. Bulletin of the Chemical Society of Japan. https://doi.org/10.1246/bcsj.20190034","ieee":"N. Zenmyo et al., “Optimized reaction pair of the CysHis tag and Ni(II)-NTA probe for highly selective chemical labeling of membrane proteins,” Bulletin of the Chemical Society of Japan, vol. 92, no. 5. Bulletin of the Chemical Society of Japan, pp. 995–1000, 2019.","ista":"Zenmyo N, Tokumaru H, Uchinomiya S, Fuchida H, Tabata S, Hamachi I, Shigemoto R, Ojida A. 2019. Optimized reaction pair of the CysHis tag and Ni(II)-NTA probe for highly selective chemical labeling of membrane proteins. Bulletin of the Chemical Society of Japan. 92(5), 995–1000.","short":"N. Zenmyo, H. Tokumaru, S. Uchinomiya, H. Fuchida, S. Tabata, I. Hamachi, R. Shigemoto, A. Ojida, Bulletin of the Chemical Society of Japan 92 (2019) 995–1000.","mla":"Zenmyo, Naoki, et al. “Optimized Reaction Pair of the CysHis Tag and Ni(II)-NTA Probe for Highly Selective Chemical Labeling of Membrane Proteins.” Bulletin of the Chemical Society of Japan, vol. 92, no. 5, Bulletin of the Chemical Society of Japan, 2019, pp. 995–1000, doi:10.1246/bcsj.20190034.","chicago":"Zenmyo, Naoki, Hiroki Tokumaru, Shohei Uchinomiya, Hirokazu Fuchida, Shigekazu Tabata, Itaru Hamachi, Ryuichi Shigemoto, and Akio Ojida. “Optimized Reaction Pair of the CysHis Tag and Ni(II)-NTA Probe for Highly Selective Chemical Labeling of Membrane Proteins.” Bulletin of the Chemical Society of Japan. Bulletin of the Chemical Society of Japan, 2019. https://doi.org/10.1246/bcsj.20190034."},"date_published":"2019-05-15T00:00:00Z","file_date_updated":"2020-10-02T08:49:58Z","ec_funded":1,"publication_status":"published","publisher":"Bulletin of the Chemical Society of Japan","department":[{"_id":"RySh"}],"acknowledgement":"his work was supported by the Grant-in-Aid for Scientific Research B (JSPS KAKENHI grant no. JP17H03090 to A. O.); the Scientific Research on Innovative Areas “Chemistry for Multimolecular Crowding Biosystems” (JSPS KAKENHI grant no. JP17H06349 to A. O.); and the European Union (European Research Council Advanced grant no. 694539 and Human Brain Project Ref. 720270 to R. S.). A. O. acknowledges the financial support of the Takeda Science Foundation.","year":"2019","date_created":"2019-07-21T21:59:16Z","date_updated":"2021-01-12T08:08:26Z","volume":92,"author":[{"first_name":"Naoki","last_name":"Zenmyo","full_name":"Zenmyo, Naoki"},{"full_name":"Tokumaru, Hiroki","last_name":"Tokumaru","first_name":"Hiroki"},{"first_name":"Shohei","last_name":"Uchinomiya","full_name":"Uchinomiya, Shohei"},{"last_name":"Fuchida","first_name":"Hirokazu","full_name":"Fuchida, Hirokazu"},{"full_name":"Tabata, Shigekazu","id":"4427179E-F248-11E8-B48F-1D18A9856A87","first_name":"Shigekazu","last_name":"Tabata"},{"last_name":"Hamachi","first_name":"Itaru","full_name":"Hamachi, Itaru"},{"full_name":"Shigemoto, Ryuichi","last_name":"Shigemoto","first_name":"Ryuichi","orcid":"0000-0001-8761-9444","id":"499F3ABC-F248-11E8-B48F-1D18A9856A87"},{"first_name":"Akio","last_name":"Ojida","full_name":"Ojida, Akio"}],"month":"05","publication_identifier":{"issn":["00092673"]},"quality_controlled":"1","project":[{"call_identifier":"H2020","name":"In situ analysis of single channel subunit composition in neurons: physiological implication in synaptic plasticity and behaviour","_id":"25CA28EA-B435-11E9-9278-68D0E5697425","grant_number":"694539"}],"oa":1,"language":[{"iso":"eng"}],"doi":"10.1246/bcsj.20190034"},{"scopus_import":"1","day":"20","article_processing_charge":"No","has_accepted_license":"1","article_type":"original","page":"256-268","publication":"iScience","citation":{"mla":"Tabata, Shigekazu, et al. “Electron Microscopic Detection of Single Membrane Proteins by a Specific Chemical Labeling.” IScience, vol. 22, no. 12, Elsevier, 2019, pp. 256–68, doi:10.1016/j.isci.2019.11.025.","short":"S. Tabata, M. Jevtic, N. Kurashige, H. Fuchida, M. Kido, K. Tani, N. Zenmyo, S. Uchinomiya, H. Harada, M. Itakura, I. Hamachi, R. Shigemoto, A. Ojida, IScience 22 (2019) 256–268.","chicago":"Tabata, Shigekazu, Marijo Jevtic, Nobutaka Kurashige, Hirokazu Fuchida, Munetsugu Kido, Kazushi Tani, Naoki Zenmyo, et al. “Electron Microscopic Detection of Single Membrane Proteins by a Specific Chemical Labeling.” IScience. Elsevier, 2019. https://doi.org/10.1016/j.isci.2019.11.025.","ama":"Tabata S, Jevtic M, Kurashige N, et al. Electron microscopic detection of single membrane proteins by a specific chemical labeling. iScience. 2019;22(12):256-268. doi:10.1016/j.isci.2019.11.025","ista":"Tabata S, Jevtic M, Kurashige N, Fuchida H, Kido M, Tani K, Zenmyo N, Uchinomiya S, Harada H, Itakura M, Hamachi I, Shigemoto R, Ojida A. 2019. Electron microscopic detection of single membrane proteins by a specific chemical labeling. iScience. 22(12), 256–268.","apa":"Tabata, S., Jevtic, M., Kurashige, N., Fuchida, H., Kido, M., Tani, K., … Ojida, A. (2019). Electron microscopic detection of single membrane proteins by a specific chemical labeling. IScience. Elsevier. https://doi.org/10.1016/j.isci.2019.11.025","ieee":"S. Tabata et al., “Electron microscopic detection of single membrane proteins by a specific chemical labeling,” iScience, vol. 22, no. 12. Elsevier, pp. 256–268, 2019."},"date_published":"2019-12-20T00:00:00Z","type":"journal_article","abstract":[{"text":"Electron microscopy (EM) is a technology that enables visualization of single proteins at a nanometer resolution. However, current protein analysis by EM mainly relies on immunolabeling with gold-particle-conjugated antibodies, which is compromised by large size of antibody, precluding precise detection of protein location in biological samples. Here, we develop a specific chemical labeling method for EM detection of proteins at single-molecular level. Rational design of α-helical peptide tag and probe structure provided a complementary reaction pair that enabled specific cysteine conjugation of the tag. The developed chemical labeling with gold-nanoparticle-conjugated probe showed significantly higher labeling efficiency and detectability of high-density clusters of tag-fused G protein-coupled receptors in freeze-fracture replicas compared with immunogold labeling. Furthermore, in ultrathin sections, the spatial resolution of the chemical labeling was significantly higher than that of antibody-mediated labeling. These results demonstrate substantial advantages of the chemical labeling approach for single protein visualization by EM.","lang":"eng"}],"issue":"12","ddc":["570"],"status":"public","title":"Electron microscopic detection of single membrane proteins by a specific chemical labeling","intvolume":" 22","user_id":"c635000d-4b10-11ee-a964-aac5a93f6ac1","_id":"7391","oa_version":"Published Version","file":[{"access_level":"open_access","file_name":"2019_iScience_Tabata.pdf","file_size":7197776,"content_type":"application/pdf","creator":"dernst","relation":"main_file","file_id":"7448","checksum":"f3e90056a49f09b205b1c4f8c739ffd1","date_created":"2020-02-04T10:48:36Z","date_updated":"2020-07-14T12:47:57Z"}],"month":"12","publication_identifier":{"issn":["2589-0042"]},"quality_controlled":"1","project":[{"name":"In situ analysis of single channel subunit composition in neurons: physiological implication in synaptic plasticity and behaviour","call_identifier":"H2020","grant_number":"694539","_id":"25CA28EA-B435-11E9-9278-68D0E5697425"},{"call_identifier":"H2020","name":"Human Brain Project Specific Grant Agreement 1 (HBP SGA 1)","_id":"25CBA828-B435-11E9-9278-68D0E5697425","grant_number":"720270"}],"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":{"pmid":["31786521"],"isi":[":000504652000020"]},"language":[{"iso":"eng"}],"doi":"10.1016/j.isci.2019.11.025","license":"https://creativecommons.org/licenses/by/4.0/","file_date_updated":"2020-07-14T12:47:57Z","ec_funded":1,"publication_status":"published","department":[{"_id":"RySh"}],"publisher":"Elsevier","year":"2019","pmid":1,"date_created":"2020-01-29T15:56:56Z","date_updated":"2024-03-28T23:30:12Z","volume":22,"author":[{"first_name":"Shigekazu","last_name":"Tabata","id":"4427179E-F248-11E8-B48F-1D18A9856A87","full_name":"Tabata, Shigekazu"},{"full_name":"Jevtic, Marijo","first_name":"Marijo","last_name":"Jevtic","id":"4BE3BC94-F248-11E8-B48F-1D18A9856A87"},{"full_name":"Kurashige, Nobutaka","first_name":"Nobutaka","last_name":"Kurashige"},{"full_name":"Fuchida, Hirokazu","first_name":"Hirokazu","last_name":"Fuchida"},{"full_name":"Kido, Munetsugu","first_name":"Munetsugu","last_name":"Kido"},{"full_name":"Tani, Kazushi","last_name":"Tani","first_name":"Kazushi"},{"first_name":"Naoki","last_name":"Zenmyo","full_name":"Zenmyo, Naoki"},{"full_name":"Uchinomiya, Shohei","last_name":"Uchinomiya","first_name":"Shohei"},{"id":"2E55CDF2-F248-11E8-B48F-1D18A9856A87","orcid":"0000-0001-7429-7896","first_name":"Harumi","last_name":"Harada","full_name":"Harada, Harumi"},{"full_name":"Itakura, Makoto","last_name":"Itakura","first_name":"Makoto"},{"first_name":"Itaru","last_name":"Hamachi","full_name":"Hamachi, Itaru"},{"full_name":"Shigemoto, Ryuichi","first_name":"Ryuichi","last_name":"Shigemoto","id":"499F3ABC-F248-11E8-B48F-1D18A9856A87","orcid":"0000-0001-8761-9444"},{"full_name":"Ojida, Akio","last_name":"Ojida","first_name":"Akio"}],"related_material":{"record":[{"status":"public","relation":"dissertation_contains","id":"11393"}]}}]