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The reporters consist of fusions of cyan\n fluorescent protein (CFP), a phosphotyrosine binding domain, a\n consensus substrate for the relevant kinase, and yellow fluorescent\n protein (YFP). Stimulation of kinase activities in living cells with\n addition of growth factors causes 20\u201335% changes in the ratios of\n yellow to cyan emissions because of phosphorylation-induced changes in\n fluorescence resonance energy transfer (FRET). Platelet-derived growth\n factor (PDGF) stimulated Abl activity most strongly in actin-rich\n membrane ruffles, supporting the importance of this tyrosine kinase in\n the regulation of cell morphology. These results establish a\n general strategy for nondestructively imaging dynamic protein tyrosine\n kinase activities with high spatial and temporal resolution in single\n living cells.<\/jats:p>","DOI":"10.1073\/pnas.211564598","type":"journal-article","created":{"date-parts":[[2002,7,26]],"date-time":"2002-07-26T10:36:44Z","timestamp":1027679804000},"page":"15003-15008","update-policy":"https:\/\/doi.org\/10.1073\/pnas.cm10313","source":"Crossref","is-referenced-by-count":394,"title":["Genetically encoded fluorescent reporters of protein tyrosine kinase activities in living cells"],"prefix":"10.1073","volume":"98","author":[{"given":"Alice Y.","family":"Ting","sequence":"first","affiliation":[{"name":"Department of Pharmacology, Howard Hughes Medical\r Institute, and Department of Chemistry and Biochemistry, University of\r California at San Diego, La Jolla, CA 92093; and\r Department of Immunology, The Scripps Research Institute,\r La Jolla, CA 92037"}]},{"given":"Kristin H.","family":"Kain","sequence":"additional","affiliation":[{"name":"Department of Pharmacology, Howard Hughes Medical\r Institute, and Department of Chemistry and Biochemistry, University of\r California at San Diego, La Jolla, CA 92093; and\r Department of Immunology, The Scripps Research Institute,\r La Jolla, CA 92037"}]},{"given":"Richard L.","family":"Klemke","sequence":"additional","affiliation":[{"name":"Department of Pharmacology, Howard Hughes Medical\r Institute, and Department of Chemistry and Biochemistry, University of\r California at San Diego, La Jolla, CA 92093; and\r Department of Immunology, The Scripps Research Institute,\r La Jolla, CA 92037"}]},{"given":"Roger Y.","family":"Tsien","sequence":"additional","affiliation":[{"name":"Department of Pharmacology, Howard Hughes Medical\r Institute, and Department of Chemistry and Biochemistry, University of\r California at San Diego, La Jolla, CA 92093; and\r Department of Immunology, The Scripps Research Institute,\r La Jolla, CA 92037"}]}],"member":"341","published-online":{"date-parts":[[2001,12,18]]},"reference":[{"key":"e_1_3_4_1_2","doi-asserted-by":"crossref","first-page":"14997","DOI":"10.1073\/pnas.211566798","volume":"98","author":"Zhang J","year":"2001","unstructured":"J Zhang, Y Ma, S S Taylor, R Y Tsien Proc Natl Acad Sci USA 98, 14997\u201315002 (2001).","journal-title":"Proc Natl Acad Sci USA"},{"key":"e_1_3_4_2_2","doi-asserted-by":"crossref","first-page":"264","DOI":"10.1016\/0076-6879(91)01025-W","volume":"201","author":"Czernik A J","year":"1991","unstructured":"A J Czernik, J-A Girault, A C Nairn, J Chen, G Snyder, J Kebabian, P Greengard Methods Enzymol 201, 264\u2013283 (1991).","journal-title":"Methods Enzymol"},{"key":"e_1_3_4_3_2","doi-asserted-by":"crossref","first-page":"1567","DOI":"10.1126\/science.290.5496.1567","volume":"290","author":"Verveer P J","year":"2000","unstructured":"P J Verveer, F S Wouters, A R Reynolds, P I Bastiaens Science 290, 1567\u20131570 (2000).","journal-title":"Science"},{"key":"e_1_3_4_4_2","doi-asserted-by":"crossref","first-page":"2085","DOI":"10.1126\/science.283.5410.2085","volume":"283","author":"Ng T","year":"1999","unstructured":"T Ng, A Squire, G Hansra, F Bornancin, C Prevostel, A Hanby, W Harris, D Barnes, S Schmidt, H Mellor, et al. 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