ATP and GTP hydrolysis assays (TLC)

Vaishnavi Rajagopal; Jon R. Lorsch

doi:10.1016/B978-0-12-420067-8.00025-8

ATP and GTP hydrolysis assays (TLC)

Vaishnavi Rajagopal, Jon R. Lorsch

School of Medicine

Research output: Chapter in Book/Report/Conference proceeding › Chapter

Abstract

Many biochemical reactions that occur within the cell are thermodynamically unfavorable. However, when these reactions are coupled to NTP (nucleoside triphosphate) hydrolysis, the energy derived from the hydrolysis of the phosphodiester bond helps drive the reaction in the favorable direction. Examples of such proteins can be found in almost all facets of cellular metabolism: glycolysis and the TCA cycle, protein biosynthesis, DNA and RNA metabolism, cellular trafficking, cell signaling, cell division etc. Thus, characterization of the NTPase activity of these proteins in vitro can help in understanding the role of the protein in complex cellular processes.

Original language	English (US)
Title of host publication	Laboratory Methods in Enzymology
Subtitle of host publication	Cell, Lipid and Carbohydrate
Publisher	Academic Press Inc.
Pages	325-334
Number of pages	10
ISBN (Print)	9780124200678
DOIs	https://doi.org/10.1016/B978-0-12-420067-8.00025-8
State	Published - Jan 1 2013

Publication series

Name	Methods in Enzymology
Volume	533
ISSN (Print)	0076-6879
ISSN (Electronic)	1557-7988

Keywords

Adenosine triphosphate (ATP) hydrolysis
Guanosine triphosphate (ATP) hydrolysis
Nucleoside diphosphate (NDP)
Nucleoside triphosphate (NTP)
Preparation of the thin-layer chromatography plate and chamber
Thin-layer chromatography (TLC) separation

ASJC Scopus subject areas

Biochemistry
Molecular Biology

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10.1016/B978-0-12-420067-8.00025-8

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title = "ATP and GTP hydrolysis assays (TLC)",

abstract = "Many biochemical reactions that occur within the cell are thermodynamically unfavorable. However, when these reactions are coupled to NTP (nucleoside triphosphate) hydrolysis, the energy derived from the hydrolysis of the phosphodiester bond helps drive the reaction in the favorable direction. Examples of such proteins can be found in almost all facets of cellular metabolism: glycolysis and the TCA cycle, protein biosynthesis, DNA and RNA metabolism, cellular trafficking, cell signaling, cell division etc. Thus, characterization of the NTPase activity of these proteins in vitro can help in understanding the role of the protein in complex cellular processes.",

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AB - Many biochemical reactions that occur within the cell are thermodynamically unfavorable. However, when these reactions are coupled to NTP (nucleoside triphosphate) hydrolysis, the energy derived from the hydrolysis of the phosphodiester bond helps drive the reaction in the favorable direction. Examples of such proteins can be found in almost all facets of cellular metabolism: glycolysis and the TCA cycle, protein biosynthesis, DNA and RNA metabolism, cellular trafficking, cell signaling, cell division etc. Thus, characterization of the NTPase activity of these proteins in vitro can help in understanding the role of the protein in complex cellular processes.

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KW - Preparation of the thin-layer chromatography plate and chamber

KW - Thin-layer chromatography (TLC) separation

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