دورية أكاديمية
Substrate Phosphorylation Rates as an In Vivo Measurement of Kinase Activity.
العنوان: | Substrate Phosphorylation Rates as an In Vivo Measurement of Kinase Activity. |
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المؤلفون: | Swaffer MP; Department of Biology, Stanford University, Stanford, CA, USA. mpswaffer@gmail.com. |
المصدر: | Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2021; Vol. 2329, pp. 19-27. |
نوع المنشور: | Journal Article |
اللغة: | English |
بيانات الدورية: | Publisher: Humana Press Country of Publication: United States NLM ID: 9214969 Publication Model: Print Cited Medium: Internet ISSN: 1940-6029 (Electronic) Linking ISSN: 10643745 NLM ISO Abbreviation: Methods Mol Biol Subsets: MEDLINE |
أسماء مطبوعة: | Publication: Totowa, NJ : Humana Press Original Publication: Clifton, N.J. : Humana Press, |
مواضيع طبية MeSH: | CDC2 Protein Kinase/*metabolism , Protein Kinase Inhibitors/*pharmacology , Schizosaccharomyces/*growth & development , Schizosaccharomyces pombe Proteins/*metabolism, CDC2 Protein Kinase/drug effects ; CDC2 Protein Kinase/genetics ; Cell Cycle/drug effects ; Fluorescence ; Microbiological Techniques ; Mutation ; Phosphorylation/drug effects ; Schizosaccharomyces/enzymology ; Schizosaccharomyces/genetics ; Schizosaccharomyces pombe Proteins/drug effects ; Schizosaccharomyces pombe Proteins/genetics ; Substrate Specificity |
مستخلص: | Measuring kinase activity in different in vivo contexts is crucial for understanding the mechanism and functions of protein kinases, such as the cyclin-dependent kinases (Cdks) and other cell cycle kinases. Here, I present the rationale and the experimental framework for an alternative approach to measure kinase activity that is based on estimating substrate phosphorylation rates in vivo. The approach presented was first developed for experiments performed to measure Cdk1 activity at different stages of the fission yeast S. pombe's cell cycle [Swaffer et al., Cell 167:1750-1761, 2016]. However, it also affords a more generalizable framework that can be adaptable to other systems and kinases, as long as specific, rapid, and reversible kinase inhibition is possible. Briefly this involves transient and reversible kinase inhibition to dephosphorylate kinase substrates in vivo, followed by quantitative measurements of phosphorylation after inhibition is removed. |
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فهرسة مساهمة: | Keywords: Cdk1; Kinase activity; Kinases; Phosphorylation |
المشرفين على المادة: | 0 (Protein Kinase Inhibitors) 0 (Schizosaccharomyces pombe Proteins) EC 2.7.11.22 (CDC2 Protein Kinase) EC 2.7.11.22 (cdc2 protein, S pombe) |
تواريخ الأحداث: | Date Created: 20210604 Date Completed: 20210824 Latest Revision: 20220113 |
رمز التحديث: | 20240628 |
DOI: | 10.1007/978-1-0716-1538-6_2 |
PMID: | 34085212 |
قاعدة البيانات: | MEDLINE |
تدمد: | 1940-6029 |
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DOI: | 10.1007/978-1-0716-1538-6_2 |