Interactions of the Rad51 Inhibitor DIDS with Human and Bovine Serum Albumins: Optical Spectroscopy and IsoThermal Calorimetry Approaches.

Fiche publication

Date publication

septembre 2019

Journal

Biochimie

Auteurs

Membres identifiés du Cancéropôle Est :
Pr NABIEV Igor

Tous les auteurs :
Velic D, Charlier C, Popova M, Jaunet-Lahary T, Bouchouireb Z, Henry S, Weigel P, Masson JY, Laurent A, Nabiev I, Fleury F

Lien Pubmed

https://www.ncbi.nlm.nih.gov/pubmed/31562898

Résumé

Rad51 is a key protein in DNA repair by homologous recombination and an important target for development of drugs in cancer therapy. 4'-diisothiocyanostilbene-2,2'-disulfonic acid (DIDS) has been used in clinic during the past 30 years as an inhibitor of anion transporters and channels. Recently DIDS has been demonstrated to affect Rad51-mediated homologous pairing and strand exchange, key processes in homologous recombination. Consequently, DIDS has been considered as a potential revertant of radio- and chemo-resistance of cancer cells, the major causes of therapy failure. Here, we have investigated the behavior of DIDS towards serum albumins. The effects of environmental factors, primarily, solvent polarity, on DIDS stability were evaluated, and the mechanisms of interaction of DIDS with human or bovine serum albumin were analyzed using isothermal calorimetry, circular dichroism and fluorescence spectroscopies. DIDS interaction with both serum albumins have been demonstrated, and the interaction characteristics have been determined. By comparing these characteristics for several DIDS derivatives, we have identified the DIDS moiety essential for the interaction. Furthermore, site competition data indicate that human albumin has two DIDS-binding sites: a high-affinity site in the IIIA subdomain and a low-affinity one in the IB subdomain. Molecular docking has revealed the key molecular moieties of DIDS responsible for its interactions in each site and shown that the IB site can bind two ligands. These findings show that binding of DIDS to serum albumin may change the balance between the free and bound DIDS forms, thereby affecting its bioavailability and efficacy against Rad51.