To perform their action, flavoproteins usually interact with a variety of low molecular weight partners, including electron transporters, yielding transient complexes whose tightness is often controlled by the redox state of the bound flavin cofactor. As a case study, here we describe the quantitative analysis of the redox-dependent interaction of the mammalian apoptosis inducing factor (AIF) with its NAD+ ligand. In particular, we report a protocol for the spectrophotometric titration of AIF in its reduced state under anaerobic conditions with NAD+, in order to determine the dissociation constant of the resulting complex.
Ligand Binding in Allosteric Flavoproteins: Part 1. Quantitative Analysis of the Interaction with NAD+ of the Apoptosis Inducing Factor (AIF) Harboring FAD in the Reduced State / P. Cocomazzi, L. Sorrentino, F. Cossu, A. Aliverti (METHODS IN MOLECULAR BIOLOGY). - In: Flavins and Flavoproteins / [a cura di] M. Barile. - Prima edizione. - New York : Springer, 2021. - ISBN 9781071612859. - pp. 179-187 [10.1007/978-1-0716-1286-6_11]
Ligand Binding in Allosteric Flavoproteins: Part 1. Quantitative Analysis of the Interaction with NAD+ of the Apoptosis Inducing Factor (AIF) Harboring FAD in the Reduced State
P. CocomazziPrimo
;F. Cossu;A. Aliverti
Ultimo
2021
Abstract
To perform their action, flavoproteins usually interact with a variety of low molecular weight partners, including electron transporters, yielding transient complexes whose tightness is often controlled by the redox state of the bound flavin cofactor. As a case study, here we describe the quantitative analysis of the redox-dependent interaction of the mammalian apoptosis inducing factor (AIF) with its NAD+ ligand. In particular, we report a protocol for the spectrophotometric titration of AIF in its reduced state under anaerobic conditions with NAD+, in order to determine the dissociation constant of the resulting complex.File | Dimensione | Formato | |
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Cocomazzi AIF anaerobic titration 2021_Protocol_LigandBindingInAllostericFlavo.pdf
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