Characterization of azido-NAD + to assess its potential as a two-dimensional infrared probe of enzyme dynamics

Samrat Dutta; Richard J Cook; Jon C.D Houtman; Amnon Kohen; Christopher M Cheatum

doi:10.1016/j.ab.2010.08.008

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Characterization of azido-NAD + to assess its potential as a two-dimensional infrared probe of enzyme dynamics

Journal article

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Characterization of azido-NAD + to assess its potential as a two-dimensional infrared probe of enzyme dynamics

Samrat Dutta, Richard J Cook, Jon C.D Houtman, Amnon Kohen and Christopher M Cheatum

Analytical biochemistry, Vol.407(2), pp.241-246

2010

DOI: 10.1016/j.ab.2010.08.008

PMID: 20705046

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https://www.ncbi.nlm.nih.gov/pmc/articles/2963086View

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Abstract

Enzyme active-site dynamics at femtosecond to picosecond time scales are of great biochemical importance, but remain relatively unexplored due to the lack of appropriate analytical methods. Two-dimensional infrared (2D IR) spectroscopy is one of the few methods that can examine chemical biological motions at this time scale, but all the IR probes used so far were specific to a few unique enzymes. The lack of IR probes of broader specificity is a major limitation to further 2D IR studies of enzyme dynamics. Here we describe the synthesis of a general IR probe for nicotinamide-dependent enzymes. This azido analog of the ubiquitous cofactor nicotinamide adenine dinucleotide is found to be stable and bind to several dehydrogenases with dissociation constants similar to that for the native cofactor. The infrared absorption spectra of this probe bound to several enzymes indicate that it has significant potential as a 2D IR probe to investigate femtosecond dynamics of nicotinamide-dependent enzymes.

Nicotinamide adenine dinucleotide

Infrared spectroscopy

Isothermal titration calorimetry

Probe

Femtosecond–picosecond

Assays

Details

Title: Subtitle: Characterization of azido-NAD + to assess its potential as a two-dimensional infrared probe of enzyme dynamics
Creators: Samrat Dutta - Department of Chemistry and Optical Science and Technology Center, University of Iowa, Iowa City, IA 52242, USA
Richard J Cook - Department of Chemistry and Optical Science and Technology Center, University of Iowa, Iowa City, IA 52242, USA
Jon C.D Houtman - Department of Microbiology, University of Iowa, Iowa City, IA 52242, USA
Amnon Kohen - Department of Chemistry and Optical Science and Technology Center, University of Iowa, Iowa City, IA 52242, USA
Christopher M Cheatum - Department of Chemistry and Optical Science and Technology Center, University of Iowa, Iowa City, IA 52242, USA
Resource Type: Journal article
Publication Details: Analytical biochemistry, Vol.407(2), pp.241-246
DOI: 10.1016/j.ab.2010.08.008
PMID: 20705046
NLM abbreviation: Anal Biochem
ISSN: 0003-2697
eISSN: 1096-0309
Publisher: Elsevier Inc
Language: English
Date published: 2010
Academic Unit: Microbiology and Immunology; Core Research Facilities; Chemistry; Internal Medicine
Record Identifier: 9983985844002771

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