{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,10]],"date-time":"2026-06-10T15:29:50Z","timestamp":1781105390767,"version":"3.54.1"},"reference-count":28,"publisher":"National Academy of Sciences","issue":"35","content-domain":{"domain":["www.pnas.org"],"crossmark-restriction":true},"short-container-title":["Proc. Natl. Acad. Sci. U.S.A."],"published-print":{"date-parts":[[2012,8,28]]},"abstract":"<jats:p>\n                    Many cellular signaling events are regulated by tyrosine phosphorylation and mediated by the opposing actions of protein tyrosine kinases and phosphatases. Protein tyrosine phosphatases are emerging as drug targets, but poor cell permeability of inhibitors has limited the development of drugs targeting these enzymes [Tautz L, et al. (2006)\n                    <jats:italic>Expert Opin Ther Targets<\/jats:italic>\n                    10:157\u2013177]. Here we developed a method to monitor tyrosine phosphatase activity at the single-cell level and applied it to the identification of cell-permeable inhibitors. The method takes advantage of the fluorogenic properties of phosphorylated coumaryl amino propionic acid (pCAP), an analog of phosphotyrosine, which can be incorporated into peptides. Once delivered into cells, pCAP peptides were dephosphorylated by protein tyrosine phosphatases, and the resulting cell fluorescence could be monitored by flow cytometry and high-content imaging. The robustness and sensitivity of the assay was validated using peptides preferentially dephosphorylated by CD45 and T-cell tyrosine phosphatase and available inhibitors of these two enzymes. The assay was applied to high-throughput screening for inhibitors of CD45, an important target for autoimmunity and infectious diseases [Hermiston ML, et al. (2003)\n                    <jats:italic>Annu Rev Immunol<\/jats:italic>\n                    21:107\u2013137]. We identified four CD45 inhibitors that showed activity in T cells and macrophages. These results indicate that our assay can be applied to primary screening for inhibitors of CD45 and of other protein tyrosine phosphatases to increase the yield of biologically active inhibitors.\n                  <\/jats:p>","DOI":"10.1073\/pnas.1205028109","type":"journal-article","created":{"date-parts":[[2012,8,13]],"date-time":"2012-08-13T22:37:49Z","timestamp":1344897469000},"page":"13972-13977","update-policy":"https:\/\/doi.org\/10.1073\/pnas.cm10313","source":"Crossref","is-referenced-by-count":31,"title":["High-throughput screen using a single-cell tyrosine phosphatase assay reveals biologically active inhibitors of tyrosine phosphatase CD45"],"prefix":"10.1073","volume":"109","author":[{"given":"Stephanie M.","family":"Stanford","sequence":"first","affiliation":[{"name":"Division of Cellular Biology, La Jolla Institute for Allergy and Immunology, La Jolla, CA 92037;"},{"name":"Institute for Genetic Medicine, University of Southern California, Los Angeles, CA 90033;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rekha G.","family":"Panchal","sequence":"additional","affiliation":[{"name":"Target Discovery &amp; Experimental Microbiology Department, Integrated Toxicology Division, United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, Frederick, MD 21702;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Logan M.","family":"Walker","sequence":"additional","affiliation":[{"name":"Division of Cellular Biology, La Jolla Institute for Allergy and Immunology, La Jolla, CA 92037;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dennis J.","family":"Wu","sequence":"additional","affiliation":[{"name":"Division of Cellular Biology, La Jolla Institute for Allergy and Immunology, La Jolla, CA 92037;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Matthew D.","family":"Falk","sequence":"additional","affiliation":[{"name":"Division of Cellular Biology, La Jolla Institute for Allergy and Immunology, La Jolla, CA 92037;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sayantan","family":"Mitra","sequence":"additional","affiliation":[{"name":"Department of Chemistry, University of Southern California, Los Angeles, CA 90089;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sagar S.","family":"Damle","sequence":"additional","affiliation":[{"name":"Division of Biology, California Institute of Technology, Pasadena, CA 91125;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"David","family":"Ruble","sequence":"additional","affiliation":[{"name":"Institute for Genetic Medicine, University of Southern California, Los Angeles, CA 90033;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Teodora","family":"Kaltcheva","sequence":"additional","affiliation":[{"name":"Department of Chemistry, University of Southern California, Los Angeles, CA 90089;"},{"name":"Department of Medicinal Chemistry, University of Utah, Salt Lake City, UT 84112; and"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sheng","family":"Zhang","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Biology, Indiana University School of Medicine, Indianapolis, IN 46202"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhong-Yin","family":"Zhang","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Biology, Indiana University School of Medicine, Indianapolis, IN 46202"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sina","family":"Bavari","sequence":"additional","affiliation":[{"name":"Target Discovery &amp; Experimental Microbiology Department, Integrated Toxicology Division, United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, Frederick, MD 21702;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Amy M.","family":"Barrios","sequence":"additional","affiliation":[{"name":"Department of Chemistry, University of Southern California, Los Angeles, CA 90089;"},{"name":"Department of Medicinal Chemistry, University of Utah, Salt Lake City, UT 84112; and"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nunzio","family":"Bottini","sequence":"additional","affiliation":[{"name":"Division of Cellular Biology, La Jolla Institute for Allergy and Immunology, La Jolla, CA 92037;"},{"name":"Institute for Genetic Medicine, University of Southern California, Los Angeles, CA 90033;"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"341","published-online":{"date-parts":[[2012,8,13]]},"reference":[{"key":"e_1_3_3_1_2","doi-asserted-by":"publisher","DOI":"10.1517\/14728222.10.1.157"},{"key":"e_1_3_3_2_2","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.immunol.21.120601.140946"},{"key":"e_1_3_3_3_2","doi-asserted-by":"publisher","DOI":"10.4155\/fmc.10.241"},{"key":"e_1_3_3_4_2","doi-asserted-by":"publisher","DOI":"10.1007\/s10555-008-9126-y"},{"key":"e_1_3_3_5_2","doi-asserted-by":"publisher","DOI":"10.2174\/092986709788612693"},{"key":"e_1_3_3_6_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.ymeth.2004.07.005"},{"key":"e_1_3_3_7_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.ymeth.2004.07.006"},{"key":"e_1_3_3_8_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2011.07.020"},{"key":"e_1_3_3_9_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.immuni.2010.03.006"},{"key":"e_1_3_3_10_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M809633200"},{"key":"e_1_3_3_11_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.bmcl.2005.08.054"},{"key":"e_1_3_3_12_2","doi-asserted-by":"publisher","DOI":"10.1021\/jm901220m"},{"key":"e_1_3_3_13_2","doi-asserted-by":"publisher","DOI":"10.1002\/cbic.200800046"},{"key":"e_1_3_3_14_2","doi-asserted-by":"publisher","DOI":"10.1042\/bj20021637"},{"key":"e_1_3_3_15_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0706233104"},{"key":"e_1_3_3_16_2","doi-asserted-by":"publisher","DOI":"10.1007\/s11095-007-9303-7"},{"key":"e_1_3_3_17_2","doi-asserted-by":"publisher","DOI":"10.1002\/cbic.200400063"},{"key":"e_1_3_3_18_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2008.11.038"},{"key":"e_1_3_3_19_2","doi-asserted-by":"publisher","DOI":"10.1084\/jem.20041890"},{"key":"e_1_3_3_20_2","doi-asserted-by":"publisher","DOI":"10.1093\/hmg\/ddn363"},{"key":"e_1_3_3_21_2","doi-asserted-by":"publisher","DOI":"10.1039\/B817853D"},{"key":"e_1_3_3_22_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.88.6.2037"},{"key":"e_1_3_3_23_2","doi-asserted-by":"publisher","DOI":"10.1021\/ja903733z"},{"key":"e_1_3_3_24_2","doi-asserted-by":"publisher","DOI":"10.1021\/jm101202j"},{"key":"e_1_3_3_25_2","first-page":"2355","article-title":"Photodynamic therapy-mediated oxidative stress can induce expression of heat shock proteins","volume":"56","author":"Gomer CJ","year":"1996","unstructured":"CJ Gomer, et al., Photodynamic therapy-mediated oxidative stress can induce expression of heat shock proteins. Cancer Res 56, 2355\u20132360 (1996).","journal-title":"Cancer Res"},{"key":"e_1_3_3_26_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.tips.2006.08.005"},{"key":"e_1_3_3_27_2","doi-asserted-by":"publisher","DOI":"10.2174\/1568026033452267"},{"key":"e_1_3_3_28_2","doi-asserted-by":"publisher","DOI":"10.1007\/978-1-60327-545-3_11"}],"container-title":["Proceedings of the National Academy of Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pnas.org\/doi\/pdf\/10.1073\/pnas.1205028109","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,4,12]],"date-time":"2022-04-12T20:57:39Z","timestamp":1649797059000},"score":1,"resource":{"primary":{"URL":"https:\/\/pnas.org\/doi\/full\/10.1073\/pnas.1205028109"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2012,8,13]]},"references-count":28,"journal-issue":{"issue":"35","published-print":{"date-parts":[[2012,8,28]]}},"alternative-id":["10.1073\/pnas.1205028109"],"URL":"https:\/\/doi.org\/10.1073\/pnas.1205028109","relation":{},"ISSN":["0027-8424","1091-6490"],"issn-type":[{"value":"0027-8424","type":"print"},{"value":"1091-6490","type":"electronic"}],"subject":[],"published":{"date-parts":[[2012,8,13]]},"assertion":[{"value":"2012-08-13","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}