An affinity capillary electrophoresis (ACE) method to estimate apparent dissociation constants between bovine brain calmodulin (CaM) and non-peptidic ligands was developed. The method was validated reproducing the dissociation constants of a number of well-known CaM ligands. In particular, the potent antagonist 125-C9 was ad hoc synthesized through an improved synthetic procedure. The ACE method was successfully applied to verify CaM affinity for lubeluzole, a well-known neuroprotective agent recently proved useful to potentiate the activity of anti-cancer drugs. Lubeluzole was slightly less potent than 125-C9 (Kd Combining double low line 2.9 ± 0.7 and 0.47 ± 0.06 1/4M, respectively) and displayed Ca2+/calmodulin-dependent kinase II (CaMKII) inhibition (IC50 Combining double low line 40 ± 1 1/4M). Possible binding modes of lubeluzole to CaM were explored by docking studies based on the X-ray crystal structures of several trifluoperazine-CaM complexes. An estimated dissociation constant in good agreement with the experimental one was found and the main aminoacidic residues and interactions contributing to complex formation were highlighted. The possibility that interference with Ca2+ pathways may contribute to the previously observed chemosensitizing effects of lubeluzole on human ovarian adenocarcinoma and lung carcinoma cells are discussed. © 2016 Elsevier Masson SAS.

The chemosensitizing agent lubeluzole binds calmodulin and inhibits Ca2+/calmodulin-dependent Kinase II

Bruno, Claudio;Cavalluzzi, Maria Maddalena
;
Lovece, Angelo;Carrieri, Antonio;Polimeno, Lorenzo;Franchini, Carlo;Lentini, Giovanni
2016-01-01

Abstract

An affinity capillary electrophoresis (ACE) method to estimate apparent dissociation constants between bovine brain calmodulin (CaM) and non-peptidic ligands was developed. The method was validated reproducing the dissociation constants of a number of well-known CaM ligands. In particular, the potent antagonist 125-C9 was ad hoc synthesized through an improved synthetic procedure. The ACE method was successfully applied to verify CaM affinity for lubeluzole, a well-known neuroprotective agent recently proved useful to potentiate the activity of anti-cancer drugs. Lubeluzole was slightly less potent than 125-C9 (Kd Combining double low line 2.9 ± 0.7 and 0.47 ± 0.06 1/4M, respectively) and displayed Ca2+/calmodulin-dependent kinase II (CaMKII) inhibition (IC50 Combining double low line 40 ± 1 1/4M). Possible binding modes of lubeluzole to CaM were explored by docking studies based on the X-ray crystal structures of several trifluoperazine-CaM complexes. An estimated dissociation constant in good agreement with the experimental one was found and the main aminoacidic residues and interactions contributing to complex formation were highlighted. The possibility that interference with Ca2+ pathways may contribute to the previously observed chemosensitizing effects of lubeluzole on human ovarian adenocarcinoma and lung carcinoma cells are discussed. © 2016 Elsevier Masson SAS.
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11586/162360
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