Sequence variation in CYP51A from the Y strain of Trypanosoma cruzi alters its sensitivity to inhibition

• Published in: FEBS letters 2014-11, Vol.588 (21), p.3878-3885
• Main Authors: Cherkesova, Tatiana S., Hargrove, Tatiana Y., Vanrell, M. Cristina, Ges, Igor, Usanov, Sergey A., Romano, Patricia S., Lepesheva, Galina I.
• Format: Article
• Language: English
• Subjects: (R)-N-(1-(2,4-dichlorophenyl)-2-(1H-1,2,4-triazol-1-yl)ethyl)-4-(5-phenyl-1,3,4-oxadiazol-2-yl)benzamide; (R)-N-(1-(2,4-dichlorophenyl)-2-(1H-imidazol-1-yl)ethyl)-4-(5-phenyl-1,3,4-oxadiazol-2-yl)benzamide; (R)-N-(1-(3,4′-difluorobiphenyl-4-yl)-2-(1H-imidazol-1-yl)ethyl)-4-(5-phenyl-1,3,4-oxadiazol-2-yl)benzamide; 14-alpha Demethylase Inhibitors - pharmacology; 14-alpha Demethylase Inhibitors - therapeutic use; Amino Acid Sequence; Amino Acid Substitution; Animals; antifungal agents; Biocatalysis; catalytic activity; Chagas disease; Chagas Disease - drug therapy; Chagas Disease - enzymology; Conserved Sequence; CYP; CYP51 sequence variation; Cytochrome P-450 Enzyme System - chemistry; Cytochrome P-450 Enzyme System - genetics; Cytochrome P-450 Enzyme System - metabolism; cytochrome P450 gene or protein; drug concentration that gives half-maximal response in cellular growth reduction; Drug resistance; drugs; EC50; GFP; green fluorescent protein; Humans; insects; Models, Molecular; Molecular Sequence Data; pathogens; proline; Protein Conformation; sequence diversity; serine; SRS; Sterol 14α-demethylase; Structure-based drug design; substrate recognition site; T. cruzi; trypanocides; Trypanosoma cruzi; Trypanosoma cruzi - drug effects; Trypanosoma cruzi - enzymology; VFV; VNI; VNT
• ISSN: 0014-5793; 1873-3468
• DOI: 10.1016/j.febslet.2014.08.030

•VNI that cures Tulahuen T. cruzi infection was found less potent against strain Y.•Amplification of CYP51 from the Y strain revealed two genes, A and B.•Y-CYP51A has a P355S substitution, which decreases its sensitivity to inhibition.•Weaker drug sensitivity of Y-CYP51A may be due to its elevated flexibility.•CYP51 structure based VNI modification produces a derivative of higher efficiency. CYP51 (sterol 14α-demethylase) is an efficient target for clinical and agricultural antifungals and an emerging target for treatment of Chagas disease, the infection that is caused by multiple strains of a protozoan pathogen Trypanosoma cruzi. Here, we analyze CYP51A from the Y strain T. cruzi. In this protein, proline 355, a residue highly conserved across the CYP51 family, is replaced with serine. The purified enzyme retains its catalytic activity, yet has been found less susceptible to inhibition. These biochemical data are consistent with cellular experiments, both in insect and human stages of the pathogen. Comparative structural analysis of CYP51 complexes with VNI and two derivatives suggests that broad-spectrum CYP51 inhibitors are likely to be preferable as antichagasic drug candidates.