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A new approach to time-resolved studies of ATP-requiring biological systems; laser flash photolysis of caged ATP
Journal article   Open access

A new approach to time-resolved studies of ATP-requiring biological systems; laser flash photolysis of caged ATP

J A McCray, L Herbette, T Kihara and D R Trentham
Proceedings of the National Academy of Sciences - PNAS, v 77(12), pp 7237-7241
01 Dec 1980
PMID: 6938971
url
https://doi.org/10.1073/pnas.77.12.7237View
Published, Version of Record (VoR)Open Access (License Unspecified) Open

Abstract

2-Nitrobenzyl derivatives have been used for several years as photolabile protecting groups in synthetic organic chemistry. Recently, P3-1-(2-nitro phenylethyladenosine 5'-triphosphate "caged ATP" was synthesized and its photolysis was shown to generate ATP in situ. This and related reactions have great potential for structural and kinetic studies of both intact and soluble biological systems and it is thus important to define the kinetic characteristics of the photolytic reaction. Caged ATP (2.5 mM) was photolyzed at 347 nm by a single 30-nsec pulse from a frequency-doubled ruby laser of 25 mJ energy to generate 500 microM ATP. The kinetics of the overall reaction were determined by monitoring the kinetics of ATP-induced dissociation of actomyosin, a reaction of known kinetic characteristics. Release of 500 microM ATP was found to be controlled by a process having a rate constant of 2.2 X 10(9) [H+] sec-1 at 22 degrees C at pH 5.8-9.5, which corresponds to 220 sec-1 at pH 7. This process is believed to be the breakdown of an aci-nitro compound, which was identified on the basis of its spectral properties and the photochromicity of related 2-nitrobenzyl compounds.

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Collaboration types
Domestic collaboration
Web of Science research areas
Biology
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