{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,29]],"date-time":"2026-10-29T22:31:38Z","timestamp":1793313098470,"version":"4.5.0"},"reference-count":32,"publisher":"American Chemical Society (ACS)","issue":"9","license":[{"start":{"date-parts":[[2020,8,18]],"date-time":"2020-08-18T00:00:00Z","timestamp":1597708800000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-029"},{"start":{"date-parts":[[2020,8,18]],"date-time":"2020-08-18T00:00:00Z","timestamp":1597708800000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-037"},{"start":{"date-parts":[[2020,8,18]],"date-time":"2020-08-18T00:00:00Z","timestamp":1597708800000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-045"}],"funder":[{"DOI":"10.13039\/501100006769","name":"Russian Science Foundation","doi-asserted-by":"publisher","award":["16-14-10364"],"award-info":[{"award-number":["16-14-10364"]}],"id":[{"id":"10.13039\/501100006769","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100006151","name":"Basic Energy Sciences","doi-asserted-by":"publisher","award":["W-31-109-Eng-38"],"award-info":[{"award-number":["W-31-109-Eng-38"]}],"id":[{"id":"10.13039\/100006151","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100012728","name":"Frederick National Laboratory for Cancer Research","doi-asserted-by":"publisher","award":["HHSN261200800001E"],"award-info":[{"award-number":["HHSN261200800001E"]}],"id":[{"id":"10.13039\/100012728","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2020,9,18]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Super-resolution fluorescent imaging in living cells remains technically challenging, largely due to the photodecomposition of fluorescent tags. The recently suggested protein-PAINT is the only super-resolution technique available for prolonged imaging of proteins in living cells. It is realized with complexes of fluorogen-activating proteins, expressed as fusions, and solvatochromic synthetic dyes. Once photobleached, the dye in the complex is replaced with a fresh fluorogen available in the sample. With suitable kinetics, this replacement creates fluorescence blinking required for attaining super-resolution and overcomes photobleaching associated with the loss of an irreplaceable fluorophore. Here we report on the rational design of two protein-PAINT tags based on the 1.58 \u00c5 crystal structure of the DiB1:M739 complex, an improved green-emitting DiB3\/F74V:M739 and a new orange-emitting DiB3\/F53L:M739. They outperform previously reported DiB-based tags to become best in class biomarkers for protein-PAINT. The new tags advance protein-PAINT from the proof-of-concept to a reliable tool suitable for prolonged super-resolution imaging of intracellular proteins in fixed and living cells and two-color PAINT-like nanoscopy with a single fluorogen.<\/jats:p>","DOI":"10.1021\/acschembio.0c00440","type":"journal-article","created":{"date-parts":[[2020,8,18]],"date-time":"2020-08-18T17:33:09Z","timestamp":1597771989000},"page":"2456-2465","source":"Crossref","is-referenced-by-count":23,"title":["Structure-Based Rational Design of Two Enhanced Bacterial\nLipocalin\n                    <i>Blc<\/i>\n                    Tags for Protein-PAINT Super-resolution\nMicroscopy"],"prefix":"10.1021","volume":"15","author":[{"given":"Liya","family":"Muslinkina","sequence":"first","affiliation":[{"name":"Frederick National Laboratory for Cancer Research , , , ,","place":["Argonne, Illinois, United States, 60439"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alexey S.","family":"Gavrikov","sequence":"additional","affiliation":[{"name":"Shemyakin\u2212Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences , ,","place":["Moscow, Russian Federation, 117997"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2164-5698","authenticated-orcid":true,"given":"Nina G.","family":"Bozhanova","sequence":"additional","affiliation":[{"name":"Vanderbilt University , , , ,","place":["Nashville, Tennessee, United States, 37235"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4935-7030","authenticated-orcid":true,"given":"Alexander S.","family":"Mishin","sequence":"additional","affiliation":[{"name":"Shemyakin\u2212Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences , ,","place":["Moscow, Russian Federation, 117997"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9339-7603","authenticated-orcid":true,"given":"Mikhail S.","family":"Baranov","sequence":"additional","affiliation":[{"name":"Shemyakin\u2212Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences , ,","place":["Moscow, Russian Federation, 117997"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jens","family":"Meiler","sequence":"additional","affiliation":[{"name":"Vanderbilt University , , , ,","place":["Nashville, Tennessee, United States, 37235"]},{"name":"Leipzig University , , , SAC ,","place":["Leipzig, Germany, 04103"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nadya V.","family":"Pletneva","sequence":"additional","affiliation":[{"name":"Shemyakin\u2212Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences , ,","place":["Moscow, Russian Federation, 117997"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vladimir Z.","family":"Pletnev","sequence":"additional","affiliation":[{"name":"Shemyakin\u2212Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences , ,","place":["Moscow, Russian Federation, 117997"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2502-4637","authenticated-orcid":true,"given":"Sergei","family":"Pletnev","sequence":"additional","affiliation":[{"name":"Frederick National Laboratory for Cancer Research , , , ,","place":["Argonne, Illinois, United States, 60439"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2020,8,18]]},"reference":[{"key":"2026081714175634700_cit1","doi-asserted-by":"crossref","first-page":"xxviii","DOI":"10.1007\/978-0-387-45524-2","volume-title":"Handbook of biological confocal microscopy","author":"Pawley","year":"2006","edition":"3"},{"issue":"11","key":"2026081714175634700_cit2","doi-asserted-by":"publisher","first-page":"3801","DOI":"10.1021\/ja044686x","article-title":"Carbocyanine\ndyes as efficient reversible single-molecule optical switch","volume":"127","author":"Heilemann","year":"2005","journal-title":"J. Am. Chem. Soc."},{"key":"2026081714175634700_cit3","doi-asserted-by":"publisher","first-page":"519","DOI":"10.1146\/annurev-physchem-032811-112012","article-title":"Live-cell super-resolution imaging with synthetic fluorophores","volume":"63","author":"van de Linde","year":"2012","journal-title":"Annu. Rev. Phys. Chem."},{"issue":"9","key":"2026081714175634700_cit4","doi-asserted-by":"publisher","first-page":"717","DOI":"10.1038\/nmeth.1489","article-title":"Live-cell super-resolution imaging\nwith trimethoprim conjugates","volume":"7","author":"Wombacher","year":"2010","journal-title":"Nat. Methods"},{"issue":"6","key":"2026081714175634700_cit5","doi-asserted-by":"publisher","first-page":"499","DOI":"10.1038\/nmeth.1605","article-title":"Fast, three-dimensional super-resolution imaging of\nlive cells","volume":"8","author":"Jones","year":"2011","journal-title":"Nat. Methods"},{"issue":"8","key":"2026081714175634700_cit6","doi-asserted-by":"publisher","first-page":"773","DOI":"10.1038\/nbt.3876","article-title":"Long time-lapse nanoscopy\nwith spontaneously blinking membrane probes","volume":"35","author":"Takakura","year":"2017","journal-title":"Nat. Biotechnol."},{"issue":"1","key":"2026081714175634700_cit7","doi-asserted-by":"publisher","first-page":"303","DOI":"10.1146\/annurev-biophys-051013-022836","article-title":"Photocontrollable\nFluorescent Proteins for Superresolution Imaging","volume":"43","author":"Shcherbakova","year":"2014","journal-title":"Annu. Rev. Biophys."},{"issue":"1","key":"2026081714175634700_cit8","doi-asserted-by":"publisher","first-page":"7","DOI":"10.1038\/nmeth0111-7b","article-title":"Live-cell dSTORM with SNAP-tag fusion proteins","volume":"8","author":"Klein","year":"2011","journal-title":"Nat. Methods"},{"issue":"8","key":"2026081714175634700_cit9","doi-asserted-by":"publisher","first-page":"681","DOI":"10.1038\/nchem.2002","article-title":"A spontaneously blinking fluorophore\nbased on intramolecular spirocyclization for live-cell super-resolution\nimaging","volume":"6","author":"Uno","year":"2014","journal-title":"Nat. Chem."},{"issue":"5","key":"2026081714175634700_cit10","doi-asserted-by":"publisher","first-page":"949","DOI":"10.1039\/C6CC09200D","article-title":"Intrinsic blinking of red fluorescent\nproteins for super-resolution microscopy","volume":"53","author":"Klementieva","year":"2017","journal-title":"Chem.\nCommun."},{"issue":"4","key":"2026081714175634700_cit11","doi-asserted-by":"publisher","first-page":"852","DOI":"10.1016\/j.bbrc.2019.11.163","article-title":"Live-cell nanoscopy with spontaneous\nblinking of conventional green fluorescent proteins","volume":"522","author":"Gavrikov","year":"2020","journal-title":"Biochem. Biophys. Res. Commun."},{"issue":"50","key":"2026081714175634700_cit12","doi-asserted-by":"publisher","first-page":"18911","DOI":"10.1073\/pnas.0609643104","article-title":"Wide-field\nsubdiffraction imaging by accumulated binding of diffusing probes","volume":"103","author":"Sharonov","year":"2006","journal-title":"Proc. Natl. Acad. Sci. U. S. A."},{"issue":"3","key":"2026081714175634700_cit13","doi-asserted-by":"publisher","first-page":"313","DOI":"10.1038\/nmeth.2835","article-title":"Multiplexed 3D cellular super-resolution\nimaging with DNA-PAINT and Exchange-PAINT","volume":"11","author":"Jungmann","year":"2014","journal-title":"Nat.\nMethods"},{"issue":"6","key":"2026081714175634700_cit14","doi-asserted-by":"publisher","first-page":"1198","DOI":"10.1038\/nprot.2017.024","article-title":"Super-resolution\nmicroscopy with DNA-PAINT","volume":"12","author":"Schnitzbauer","year":"2017","journal-title":"Nat. Protoc."},{"key":"2026081714175634700_cit15","doi-asserted-by":"publisher","first-page":"95","DOI":"10.1007\/978-1-62703-137-0_7","article-title":"High-content\nsuper-resolution imaging of live cell by uPAINT","volume":"950","author":"Giannone","year":"2013","journal-title":"Methods Mol. Biol. (N. Y., NY, U. S.)"},{"issue":"10","key":"2026081714175634700_cit16","doi-asserted-by":"publisher","first-page":"7138","DOI":"10.1039\/C7SC01628J","article-title":"Protein\nlabeling for live cell fluorescence microscopy with a highly photostable\nrenewable signal","volume":"8","author":"Bozhanova","year":"2017","journal-title":"Chem. Sci."},{"issue":"41","key":"2026081714175634700_cit17","doi-asserted-by":"publisher","first-page":"13234","DOI":"10.1002\/chem.201403678","article-title":"Red-shifted\nfluorescent aminated derivatives of a conformationally locked GFP\nchromophore","volume":"20","author":"Baranov","year":"2014","journal-title":"Chem. - Eur. J."},{"issue":"3","key":"2026081714175634700_cit18","doi-asserted-by":"publisher","first-page":"233","DOI":"10.1007\/s40259-018-0278-1","article-title":"Anticalin\u00ad((R)) Proteins\nas Therapeutic Agents in Human Diseases","volume":"32","author":"Rothe","year":"2018","journal-title":"BioDrugs"},{"issue":"1\u20133","key":"2026081714175634700_cit19","doi-asserted-by":"publisher","first-page":"183","DOI":"10.1016\/S0014-5793(04)00199-1","article-title":"The crystal structure of the Escherichia coli lipocalin\nBlc suggests a possible role in phospholipid binding","volume":"562","author":"Campanacci","year":"2004","journal-title":"FEBS Lett."},{"issue":"20","key":"2026081714175634700_cit20","doi-asserted-by":"publisher","first-page":"4877","DOI":"10.1016\/j.febslet.2006.07.086","article-title":"The membrane bound bacterial lipocalin\nBlc is a functional dimer with binding preference for lysophospholipids","volume":"580","author":"Campanacci","year":"2006","journal-title":"FEBS Lett."},{"issue":"12","key":"2026081714175634700_cit21","doi-asserted-by":"publisher","first-page":"1308","DOI":"10.1107\/S0907444910039375","article-title":"Structural and biochemical analyses reveal a monomeric\nstate of the bacterial lipocalin Blc","volume":"66","author":"Schiefner","year":"2010","journal-title":"Acta Crystallogr.,\nSect. D: Biol. Crystallogr."},{"issue":"1","key":"2026081714175634700_cit22","doi-asserted-by":"publisher","first-page":"121","DOI":"10.1002\/prot.10497","article-title":"Crystallographic\nanalysis of an \u201canticalin\u201d with tailored specificity\nfor fluorescein reveals high structural plasticity of the lipocalin\nloop region","volume":"53","author":"Korndorfer","year":"2003","journal-title":"Proteins: Struct., Funct., Genet."},{"issue":"3","key":"2026081714175634700_cit23","doi-asserted-by":"publisher","first-page":"499","DOI":"10.1016\/0022-2836(87)90296-8","article-title":"Molecular\nstructure of the bilin binding protein (BBP) from Pieris brassicae\nafter refinement at 2.0 A resolution","volume":"198","author":"Huber","year":"1987","journal-title":"J. Mol.\nBiol."},{"issue":"41","key":"2026081714175634700_cit24","doi-asserted-by":"publisher","first-page":"11513","DOI":"10.1073\/pnas.1525622113","article-title":"Blue protein\nwith red fluorescence","volume":"113","author":"Ghosh","year":"2016","journal-title":"Proc. Natl. Acad. Sci.\nU. S. A."},{"issue":"7","key":"2026081714175634700_cit25","doi-asserted-by":"publisher","first-page":"1602","DOI":"10.1016\/j.cell.2013.05.038","article-title":"A bilirubin-inducible\nfluorescent protein from eel muscle","volume":"153","author":"Kumagai","year":"2013","journal-title":"Cell"},{"issue":"3","key":"2026081714175634700_cit26","doi-asserted-by":"publisher","first-page":"351","DOI":"10.1515\/hsz-2018-0342","article-title":"An engineered lipocalin that tightly complexes the\nplant poison colchicine for use as antidote and in bioanalytical applications","volume":"400","author":"Barkovskiy","year":"2019","journal-title":"Biol. Chem."},{"issue":"6","key":"2026081714175634700_cit27","doi-asserted-by":"publisher","first-page":"1405","DOI":"10.1111\/j.1751-1097.2007.00180.x","article-title":"Characterization of fluorescence\nof ANS-tear lipocalin complex: evidence for multiple-binding modes","volume":"83","author":"Gasymov","year":"2007","journal-title":"Photochem. Photobiol."},{"issue":"3","key":"2026081714175634700_cit28","doi-asserted-by":"publisher","first-page":"872","DOI":"10.1016\/j.bbrc.2018.01.134","article-title":"A dual-ligand-modulable fluorescent\nprotein based on UnaG and calmodulin","volume":"496","author":"Shitashima","year":"2018","journal-title":"Biochem.\nBiophys. Res. Commun."},{"key":"2026081714175634700_cit29","doi-asserted-by":"publisher","first-page":"15348","DOI":"10.1038\/srep15348","article-title":"Light-induced cell damage in live-cell\nsuper-resolution microscopy","volume":"5","author":"Waldchen","year":"2015","journal-title":"Sci. Rep."},{"issue":"6","key":"2026081714175634700_cit30","doi-asserted-by":"publisher","first-page":"1302","DOI":"10.1016\/j.cell.2016.02.058","article-title":"SnapShot: Timescales in Cell Biology","volume":"164","author":"Shamir","year":"2016","journal-title":"Cell"},{"issue":"6","key":"2026081714175634700_cit31","doi-asserted-by":"publisher","first-page":"1115","DOI":"10.1021\/acschembio.9b00149","article-title":"Single-Molecule\nLocalization Microscopy with the Fluorescence-Activating and Absorption-Shifting\nTag (FAST) System","volume":"14","author":"Smith","year":"2019","journal-title":"ACS Chem. Biol."},{"issue":"2","key":"2026081714175634700_cit32","doi-asserted-by":"publisher","first-page":"758","DOI":"10.1021\/acs.chemrev.6b00238","article-title":"Photoinduced Chemistry in Fluorescent\nProteins: Curse or Blessing?","volume":"117","author":"Acharya","year":"2017","journal-title":"Chem. Rev."}],"container-title":["ACS Chemical Biology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pubs.acs.org\/doi\/pdf\/10.1021\/acschembio.0c00440","content-type":"application\/pdf","content-version":"vor","intended-application":"unspecified"},{"URL":"https:\/\/pubs.acs.org\/acbcct\/article-pdf\/15\/9\/2456\/9225257\/cb0c00440.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/pubs.acs.org\/acbcct\/article-pdf\/15\/9\/2456\/9225257\/cb0c00440.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,8,17]],"date-time":"2026-08-17T18:27:38Z","timestamp":1786991258000},"score":1,"resource":{"primary":{"URL":"https:\/\/pubs.acs.org\/acbcct\/article\/15\/9\/2456\/865936\/Structure-Based-Rational-Design-of-Two-Enhanced"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,8,18]]},"references-count":32,"journal-issue":{"issue":"9","published-online":{"date-parts":[[2020,8,18]]},"published-print":{"date-parts":[[2020,9,18]]}},"URL":"https:\/\/doi.org\/10.1021\/acschembio.0c00440","relation":{},"ISSN":["1554-8929","1554-8937"],"issn-type":[{"value":"1554-8929","type":"print"},{"value":"1554-8937","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,8,18]]}}}