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Successful labeling of proteins on the cell membrane has been performed using mutant \u03b2\u2010lactamase tag (BL\u2010tag) technology. In the present study, intracellular protein labeling with novel cell membrane permeable probes based on \u03b2\u2010lactam prodrugs is described. The prodrug\u2010based probes quickly permeated the plasma membranes of living mammalian cells, and efficiently labeled intracellular proteins at low probe concentrations. Because these cell\u2010permeable probes were activated only inside cells, simultaneous discriminative labeling of intracellular and cell surface BL\u2010tag fusion proteins was attained by using cell\u2010permeable and impermeable probes. Thus, this technology enables adequate discrimination of the location of proteins labeled with the same protein tag, in conjunction with different color probes, by dual\u2010color fluorescence. Moreover, the combination of BL\u2010tag technology and the prodrug\u2010based probes enabled the labeling of target proteins without requiring a washing step, owing to the efficient entry of probes into cells and the fast covalent labeling achieved with BL\u2010tag technology after bioactivation. This prodrug\u2010based probe design strategy for BL\u2010tags provides a simple experimental procedure with application to cellular studies with the additional advantage of reduced stress to living cells.<\/jats:p>","DOI":"10.1002\/chem.201100973","type":"journal-article","created":{"date-parts":[[2011,6,8]],"date-time":"2011-06-08T08:52:16Z","timestamp":1307523136000},"page":"8342-8349","source":"Crossref","is-referenced-by-count":30,"title":["Intracellular Protein Labeling with Prodrug\u2010Like Probes Using a Mutant \u03b2\u2010Lactamase Tag"],"prefix":"10.1002","volume":"17","author":[{"given":"Shuji","family":"Watanabe","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shin","family":"Mizukami","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuri","family":"Akimoto","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuichiro","family":"Hori","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kazuya","family":"Kikuchi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2011,6,8]]},"reference":[{"key":"e_1_2_6_1_2","unstructured":"\u00a0"},{"key":"e_1_2_6_2_2","doi-asserted-by":"publisher","DOI":"10.1002\/cbic.200700158"},{"key":"e_1_2_6_3_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.sbi.2007.07.005"},{"key":"e_1_2_6_4_2","unstructured":"\u00a0"},{"key":"e_1_2_6_5_2","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth735"},{"key":"e_1_2_6_6_2","doi-asserted-by":"publisher","DOI":"10.1021\/ja0604111"},{"key":"e_1_2_6_7_2","doi-asserted-by":"publisher","DOI":"10.1021\/ja047749k"},{"key":"e_1_2_6_8_2","doi-asserted-by":"publisher","DOI":"10.1021\/ja048396s"},{"key":"e_1_2_6_9_2","doi-asserted-by":"publisher","DOI":"10.1021\/ja039915e"},{"key":"e_1_2_6_10_2","doi-asserted-by":"publisher","DOI":"10.1002\/cbic.200700614"},{"key":"e_1_2_6_11_2","unstructured":"\u00a0"},{"key":"e_1_2_6_12_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.281.5374.269"},{"key":"e_1_2_6_13_2","doi-asserted-by":"publisher","DOI":"10.1021\/ja017687n"},{"key":"e_1_2_6_14_2","doi-asserted-by":"publisher","DOI":"10.1021\/ja910703v"},{"key":"e_1_2_6_15_2","doi-asserted-by":"publisher","DOI":"10.1038\/nbt954"},{"key":"e_1_2_6_16_2","doi-asserted-by":"publisher","DOI":"10.1021\/ja807872s"},{"key":"e_1_2_6_17_2","unstructured":"\u00a0"},{"key":"e_1_2_6_18_2","doi-asserted-by":"publisher","DOI":"10.1038\/nbt765"},{"key":"e_1_2_6_19_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0401923101"},{"key":"e_1_2_6_20_2","doi-asserted-by":"publisher","DOI":"10.1021\/cb800025k"},{"key":"e_1_2_6_21_2","doi-asserted-by":"publisher","DOI":"10.1002\/ange.200352852"},{"key":"e_1_2_6_21_3","doi-asserted-by":"publisher","DOI":"10.1002\/anie.200352852"},{"key":"e_1_2_6_22_2","doi-asserted-by":"publisher","DOI":"10.1021\/ja904800k"},{"key":"e_1_2_6_23_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.copbio.2010.09.011"},{"key":"e_1_2_6_24_2","unstructured":"\u00a0"},{"key":"e_1_2_6_25_2","doi-asserted-by":"publisher","DOI":"10.1021\/ja8082285"},{"key":"e_1_2_6_26_2","doi-asserted-by":"publisher","DOI":"10.1021\/bc100333k"},{"key":"e_1_2_6_27_2","doi-asserted-by":"publisher","DOI":"10.1039\/c0cc02432e"},{"key":"e_1_2_6_28_2","first-page":"853","volume":"266","author":"Christensen H.","year":"1990","journal-title":"Biochem. 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