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Most photoactivatable fluorophores are relatively large in size and need to be activated by ultraviolet light; this dramatically limits their applications. To introduce photoactivatable fluorophores into proteins, recent investigations have explored several protein\u2010labeling technologies, including fluorescein arsenical hairpin (FlAsH) Tag, HaloTag labeling, SNAPTag labeling, and other bioorthogonal chemistry\u2010based methods. However, these technologies require a multistep labeling process. Here, by using genetic code expansion and a single sulfur\u2010for\u2010oxygen atom replacement within an existing fluorescent amino acid, we have site\u2010specifically incorporated the photoactivatable fluorescent amino acid thioacridonylalanine (SAcd) into proteins in a single step. Moreover, upon exposure to visible light, SAcd can be efficiently desulfurized to its oxo derivatives, thus restoring the strong fluorescence of labeled proteins.<\/jats:p>","DOI":"10.1002\/cbic.202000602","type":"journal-article","created":{"date-parts":[[2020,9,22]],"date-time":"2020-09-22T20:54:29Z","timestamp":1600808069000},"page":"501-504","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":14,"title":["Site\u2010Specific Incorporation of a Photoactivatable Fluorescent Amino Acid"],"prefix":"10.1002","volume":"22","author":[{"given":"Juan","family":"Tang","sequence":"first","affiliation":[{"name":"Department of Chemistry Rice University  6100 Main Street Houston TX 77005 USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chenfei","family":"Yu","sequence":"additional","affiliation":[{"name":"Department of Chemistry Rice University  6100 Main Street Houston TX 77005 USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Axel","family":"Loredo","sequence":"additional","affiliation":[{"name":"Department of Chemistry Rice University  6100 Main Street Houston TX 77005 USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuda","family":"Chen","sequence":"additional","affiliation":[{"name":"Department of Chemistry Rice University  6100 Main Street Houston TX 77005 USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4311-971X","authenticated-orcid":false,"given":"Han","family":"Xiao","sequence":"additional","affiliation":[{"name":"Department of Chemistry Rice University  6100 Main Street Houston TX 77005 USA"},{"name":"Department of Biosciences Rice University  6100 Main Street Houston TX 77005 USA"},{"name":"Department of Bioengineering Rice University  6100 Main Street Houston TX 77005 USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2020,11,2]]},"reference":[{"key":"e_1_2_2_1_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bmc.2010.07.011"},{"key":"e_1_2_2_2_1","doi-asserted-by":"publisher","DOI":"10.1002\/anie.201202134"},{"key":"e_1_2_2_3_1","doi-asserted-by":"publisher","DOI":"10.1039\/c2pp05342j"},{"key":"e_1_2_2_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.febslet.2014.06.043"},{"key":"e_1_2_2_5_1","doi-asserted-by":"publisher","DOI":"10.1021\/cb600132m"},{"key":"e_1_2_2_6_1","first-page":"1","volume-title":"Prog. 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