Optimized Fluorescent Trimethoprim Derivatives for in vivo Protein Labeling Export

@article{citeulike:1512140, abstract = {The combined technologies of optical microscopy and selective probes allow for real-time analysis of protein function in living cells. Synthetic chemistry offers a means to develop specific, protein-targeted probes that exhibit greater optical and chemical functionality than the widely used fluorescent proteins. Here we describe pharmacokinetically optimized, fluorescent trimethoprim (TMP) analogues that can be used to specifically label recombinant proteins fused to E. coli dihydrofolate reductase (eDHFR) in living, wild-type mammalian cells. These improved fluorescent tags exhibited high specificity and fast labeling kinetics, and they could be detected at a high signal-to-noise ratio by using fluorescence microscopy and fluorescence-activated cell sorting (FACS). We also show that fluorescent TMP-eDHFR complexes are complements to green fluorescent protein (GFP) for two-color protein labeling experiments in cells.}, address = {Department of Chemistry and Chemical Biology, Cornell University, Ithaca, NY 14853, USA; Active Motif, Inc. 1914 Palomar Oaks Way, Suite 150, Carlsbad, CA 92008, USA; Department of Biological Sciences, Columbia University, 1244 Amsterdam Avenue, New York, NY 10027, USA; Department of Chemistry, University of Illinois at Chicago, 845 W. Taylor St., MC 111, Chicago, IL 60607, USA, Fax: (+1) 312-996-0431; Department of Chemistry, Columbia University, 3000 Broadway, MC 3111, New York, NY 10027, USA, Fax: (+1)212-932-1289}, author = {Calloway, Nathaniel T. and Choob, Michael and Sanz, Ana and Sheetz, Michael P. and Miller, Lawrence W. and Cornish, Virginia W.}, citeulike-article-id = {1512140}, citeulike-linkout-0 = {http://dx.doi.org/10.1002/cbic.200600414}, citeulike-linkout-1 = {http://view.ncbi.nlm.nih.gov/pubmed/17378009}, citeulike-linkout-2 = {http://www.hubmed.org/display.cgi?uids=17378009}, citeulike-linkout-3 = {http://www3.interscience.wiley.com/cgi-bin/abstract/114194225/ABSTRACT}, doi = {10.1002/cbic.200600414}, journal = {ChemBioChem}, keywords = {chemistry, dhfr, labelling, protein}, number = {7}, pages = {767--774}, posted-at = {2007-08-14 09:18:00}, priority = {3}, title = {Optimized Fluorescent Trimethoprim Derivatives for in vivo Protein Labeling}, url = {http://dx.doi.org/10.1002/cbic.200600414}, volume = {8}, year = {2007} }

TY - JOUR ID - citeulike:1512140 L3 - citeulike-article-id:1512140 N2 - The combined technologies of optical microscopy and selective probes allow for real-time analysis of protein function in living cells. Synthetic chemistry offers a means to develop specific, protein-targeted probes that exhibit greater optical and chemical functionality than the widely used fluorescent proteins. Here we describe pharmacokinetically optimized, fluorescent trimethoprim (TMP) analogues that can be used to specifically label recombinant proteins fused to E. coli dihydrofolate reductase (eDHFR) in living, wild-type mammalian cells. These improved fluorescent tags exhibited high specificity and fast labeling kinetics, and they could be detected at a high signal-to-noise ratio by using fluorescence microscopy and fluorescence-activated cell sorting (FACS). We also show that fluorescent TMP-eDHFR complexes are complements to green fluorescent protein (GFP) for two-color protein labeling experiments in cells. IS - 7 JF - ChemBioChem AD - Department of Chemistry and Chemical Biology, Cornell University, Ithaca, NY 14853, USA; Active Motif, Inc. 1914 Palomar Oaks Way, Suite 150, Carlsbad, CA 92008, USA; Department of Biological Sciences, Columbia University, 1244 Amsterdam Avenue, New York, NY 10027, USA; Department of Chemistry, University of Illinois at Chicago, 845 W. Taylor St., MC 111, Chicago, IL 60607, USA, Fax: (+1) 312-996-0431; Department of Chemistry, Columbia University, 3000 Broadway, MC 3111, New York, NY 10027, USA, Fax: (+1)212-932-1289 EP - 774 TI - Optimized Fluorescent Trimethoprim Derivatives for in vivo Protein Labeling VL - 8 SP - 767 KW - chemistry KW - dhfr KW - labelling KW - protein AU - Calloway, Nathaniel AU - Choob, Michael AU - Sanz, Ana AU - Sheetz, Michael AU - Miller, Lawrence AU - Cornish, Virginia PY - 2007/// UR - http://dx.doi.org/10.1002/cbic.200600414 ER -