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. Cocomazzi
Primo
;
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.
Protein–ligand interaction; Dissociation constant; Electron carrier; Charge-transfer complex; Anaerobiosis; Photoreduction; Spectrophotometric titration
Settore BIO/10 - Biochimica
Book Part (author)
File in questo prodotto:
File Dimensione Formato  
Cocomazzi AIF anaerobic titration 2021_Protocol_LigandBindingInAllostericFlavo.pdf

accesso riservato

Tipologia: Publisher's version/PDF
Dimensione 278.77 kB
Formato Adobe PDF
278.77 kB Adobe PDF   Visualizza/Apri   Richiedi una copia
Pubblicazioni consigliate

I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.

Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2434/829089
Citazioni
  • ???jsp.display-item.citation.pmc??? 0
  • Scopus 1
  • ???jsp.display-item.citation.isi??? 1
social impact