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  Nanopore Enzymology to Study Protein Kinases and Their Inhibition by Small Molecules.

Harrington, L., Alexander, L. T., Knapp, S., & Bayley, H. (2021). Nanopore Enzymology to Study Protein Kinases and Their Inhibition by Small Molecules. In M. A. Fahie (Ed.), Nanopore Technology. Methods in Molecular Biology, vol 2186. (pp. 95-114). New York: Humana. doi:10.1007/978-1-0716-0806-7_8.

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 Creators:
Harrington, Leon1, Author              
Alexander, Leila T2, Author
Knapp, Stefan2, Author
Bayley, Hagan2, Author
Affiliations:
1Schwille, Petra / Cellular and Molecular Biophysics, Max Planck Institute of Biochemistry, Max Planck Society, ou_1565169              
2external, ou_persistent22              

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Free keywords: Enzyme kinetics; Enzyme mechanism; Inhibitor screening; Kinase; Kinase inhibitors; Nanopore enzymology; Single molecule
 Abstract: Nanopore enzymology is a powerful single-molecule technique for the label-free study of enzymes using engineered protein nanopore sensors. The technique has been applied to protein kinases, where it has enabled the full repertoire of kinase function to be observed, including: kinetics of substrate binding and dissociation, product binding and dissociation, nucleotide binding, and reversible phosphorylation. Further, minor modifications enable the screening of type I kinase inhibitors and the determination of inhibition constants in a facile and label-free manner. Here, we describe the design and production of suitably engineered protein nanopores and their use for the determination of key mechanistic parameters of kinases. We also provide procedures for the determination of inhibition constants of protein kinase inhibitors.

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Language(s): eng - English
 Dates: 2020-092021
 Publication Status: Published in print
 Pages: -
 Publishing info: -
 Table of Contents: -
 Rev. Type: -
 Identifiers: ISI: 32918732
DOI: 10.1007/978-1-0716-0806-7_8
 Degree: -

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Title: Nanopore Technology. Methods in Molecular Biology, vol 2186.
Source Genre: Book
 Creator(s):
Fahie, M. A., Editor
Affiliations:
-
Publ. Info: New York : Humana
Pages: - Volume / Issue: 2186 Sequence Number: - Start / End Page: 95 - 114 Identifier: ISBN: 978-1-0716-0805-0