Interrogating Signaling Nodes Involved in Cellular Transformations Using Kinase Activity Probes
Protein kinases catalyze protein phosphorylation and thereby control the flow of information through signaling cascades. Currently available methods for concomitant assessment of the enzymatic activities of multiple kinases in complex biological samples rely on indirect proxies for enzymatic activit...
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| Format: | Article |
| Language: | en_US |
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Elsevier
2014
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| Online Access: | http://hdl.handle.net/1721.1/91552 https://orcid.org/0000-0002-5749-7869 https://orcid.org/0000-0002-1801-5548 |
| _version_ | 1826198143110217728 |
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| author | Stains, Cliff I. Tedford, Nathan C. Walkup, Traci C. Lukovic, Elvedin Goguen, Brenda N. Griffith, Linda G. Lauffenburger, Douglas A. Imperiali, Barbara |
| author2 | Massachusetts Institute of Technology. Department of Biological Engineering |
| author_facet | Massachusetts Institute of Technology. Department of Biological Engineering Stains, Cliff I. Tedford, Nathan C. Walkup, Traci C. Lukovic, Elvedin Goguen, Brenda N. Griffith, Linda G. Lauffenburger, Douglas A. Imperiali, Barbara |
| author_sort | Stains, Cliff I. |
| collection | MIT |
| description | Protein kinases catalyze protein phosphorylation and thereby control the flow of information through signaling cascades. Currently available methods for concomitant assessment of the enzymatic activities of multiple kinases in complex biological samples rely on indirect proxies for enzymatic activity, such as posttranslational modifications to protein kinases. Our laboratories have recently described a method for directly quantifying the enzymatic activity of kinases in unfractionated cell lysates using substrates containing a phosphorylation-sensitive unnatural amino acid termed CSox, which can be monitored using fluorescence. Here, we demonstrate the utility of this method using a probe set encompassing p38α, MK2, ERK1/2, Akt, and PKA. This panel of chemosensors provides activity measurements of individual kinases in a model of skeletal muscle differentiation and can be readily used to generate individualized kinase activity profiles for tissue samples from clinical cancer patients. |
| first_indexed | 2024-09-23T10:59:41Z |
| format | Article |
| id | mit-1721.1/91552 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T10:59:41Z |
| publishDate | 2014 |
| publisher | Elsevier |
| record_format | dspace |
| spelling | mit-1721.1/915522022-09-27T16:23:38Z Interrogating Signaling Nodes Involved in Cellular Transformations Using Kinase Activity Probes Stains, Cliff I. Tedford, Nathan C. Walkup, Traci C. Lukovic, Elvedin Goguen, Brenda N. Griffith, Linda G. Lauffenburger, Douglas A. Imperiali, Barbara Massachusetts Institute of Technology. Department of Biological Engineering Massachusetts Institute of Technology. Department of Biology Massachusetts Institute of Technology. Department of Chemistry Stains, Cliff I. Tedford, Nathan C. Walkup, Traci C. Lukovic, Elvedin Goguen, Brenda N. Griffith, Linda G. Lauffenburger, Douglas A. Imperiali, Barbara Protein kinases catalyze protein phosphorylation and thereby control the flow of information through signaling cascades. Currently available methods for concomitant assessment of the enzymatic activities of multiple kinases in complex biological samples rely on indirect proxies for enzymatic activity, such as posttranslational modifications to protein kinases. Our laboratories have recently described a method for directly quantifying the enzymatic activity of kinases in unfractionated cell lysates using substrates containing a phosphorylation-sensitive unnatural amino acid termed CSox, which can be monitored using fluorescence. Here, we demonstrate the utility of this method using a probe set encompassing p38α, MK2, ERK1/2, Akt, and PKA. This panel of chemosensors provides activity measurements of individual kinases in a model of skeletal muscle differentiation and can be readily used to generate individualized kinase activity profiles for tissue samples from clinical cancer patients. Cell Migration Consortium National Institutes of Health (U.S.) (GM064346) National Institutes of Health (U.S.). Tumor Cell Networks Center (U54-CA112967) National Science Foundation (U.S.) (NSF-0070319) National Institutes of Health (U.S.). Ruth L. Kirschstein National Research Service Award (Fellowship F32GM085909) 2014-11-13T19:25:03Z 2014-11-13T19:25:03Z 2012-02 2011-10 Article http://purl.org/eprint/type/JournalArticle 10745521 http://hdl.handle.net/1721.1/91552 Stains, Cliff I., Nathan C. Tedford, Traci C. Walkup, Elvedin Lukovic, Brenda N. Goguen, Linda G. Griffith, Douglas A. Lauffenburger, and Barbara Imperiali. “Interrogating Signaling Nodes Involved in Cellular Transformations Using Kinase Activity Probes.” Chemistry & Biology 19, no. 2 (February 2012): 210–217. © 2012 Elsevier Ltd. https://orcid.org/0000-0002-5749-7869 https://orcid.org/0000-0002-1801-5548 en_US http://dx.doi.org/10.1016/j.chembiol.2011.11.012 Chemistry and Biology Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use. application/pdf Elsevier Elsevier |
| spellingShingle | Stains, Cliff I. Tedford, Nathan C. Walkup, Traci C. Lukovic, Elvedin Goguen, Brenda N. Griffith, Linda G. Lauffenburger, Douglas A. Imperiali, Barbara Interrogating Signaling Nodes Involved in Cellular Transformations Using Kinase Activity Probes |
| title | Interrogating Signaling Nodes Involved in Cellular Transformations Using Kinase Activity Probes |
| title_full | Interrogating Signaling Nodes Involved in Cellular Transformations Using Kinase Activity Probes |
| title_fullStr | Interrogating Signaling Nodes Involved in Cellular Transformations Using Kinase Activity Probes |
| title_full_unstemmed | Interrogating Signaling Nodes Involved in Cellular Transformations Using Kinase Activity Probes |
| title_short | Interrogating Signaling Nodes Involved in Cellular Transformations Using Kinase Activity Probes |
| title_sort | interrogating signaling nodes involved in cellular transformations using kinase activity probes |
| url | http://hdl.handle.net/1721.1/91552 https://orcid.org/0000-0002-5749-7869 https://orcid.org/0000-0002-1801-5548 |
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