In silico and in vitro analysis of genetic variants of the equine CYP3A94, CYP3A95 and CYP3A97 isoenzymes.

Vimercati, Sara; Elli, S; Jagannathan, Vidya; Pandey, Amit Vikram; Peduto, Nadja; Leeb, Tosso; Mevissen, Meike (2019). In silico and in vitro analysis of genetic variants of the equine CYP3A94, CYP3A95 and CYP3A97 isoenzymes. Toxicology in vitro, 60, pp. 116-124. Elsevier 10.1016/j.tiv.2019.05.011

[img] Text
Vimercati_2019_Toxicol_in_vitro_60_116_124.pdf - Published Version
Restricted to registered users only
Available under License Publisher holds Copyright.

Download (2MB) | Request a copy

Cytochrome P450 enzymes (CYPs) of the equine CYP3A subfamily are predominantly involved in drug metabolism. In this study, genetic variants of the equine CYP3A94, CYP3A95, and CYP3A97 were identified and characterized using in silico modeling and in vitro enzyme kinetics. The genomes of 81 horses were sequenced to obtain the genetic variants. Structural CYP modifications of the most frequent variants were analyzed in silico using the 3D-structures predicted by homology modeling. Enzyme kinetic analyses were performed using testosterone as substrate. Twenty genetic variants were found including five missense variants (CYP3A94:p.Asp217Asn, CYP3A95:p.Asp214His, CYP3A95:p.Ser392Thr, CYP3A97:p.Ile119Thr, CYP3A97:p.Met500Val) with a higher percentage of minor allele frequency (MAF) (range 0.2-0.4). A splice-site variant (c.798 + 1G > A) in CYP3A94, likely to generate a truncated protein, was found in 50% of the horses. CYP3A94:p.Asp217Asn and CYP3A95:p.Asp214His were localized on the CYP F-α-helix, an important region for the substrate interactions in the human CYP3A4. Testosterone 2β-hydroxylation was diminished in CYP3A94Asn and CYP3A95Thr. Ketoconazole inhibited 2β-hydroxylation differently in the five variants with the most pronounced inhibition obtained for CYP3A95Thr. In vitro and in silico analyses of genetic variants allow unraveling structural features in equine CYPs that correlate with changes in the CYP activity.

Item Type:

Journal Article (Original Article)

Division/Institute:

04 Faculty of Medicine > Department of Gynaecology, Paediatrics and Endocrinology (DFKE) > Clinic of Paediatric Medicine
05 Veterinary Medicine > Department of Clinical Research and Veterinary Public Health (DCR-VPH) > Veterinary Pharmacology and Toxicology
05 Veterinary Medicine > Department of Clinical Research and Veterinary Public Health (DCR-VPH) > Institute of Genetics
04 Faculty of Medicine > Pre-clinic Human Medicine > BioMedical Research (DBMR) > Unit Childrens Hospital
05 Veterinary Medicine > Department of Clinical Research and Veterinary Public Health (DCR-VPH)

Graduate School:

Graduate School for Cellular and Biomedical Sciences (GCB)

UniBE Contributor:

Vimercati, Sara; Jagannathan, Vidya; Pandey, Amit Vikram; Peduto, Nadja; Leeb, Tosso and Mevissen, Meike

Subjects:

600 Technology > 610 Medicine & health
500 Science > 570 Life sciences; biology
500 Science > 590 Animals (Zoology)
600 Technology > 630 Agriculture

ISSN:

0887-2333

Publisher:

Elsevier

Language:

English

Submitter:

Tosso Leeb

Date Deposited:

06 Jun 2019 09:26

Last Modified:

24 Oct 2019 12:05

Publisher DOI:

10.1016/j.tiv.2019.05.011

PubMed ID:

31108125

Uncontrolled Keywords:

Cytochrome P450 enzymes (CYPs) Drug metabolism Equine CYPs Genetic variants Homology modeling

BORIS DOI:

10.7892/boris.130816

URI:

https://boris.unibe.ch/id/eprint/130816

Actions (login required)

Edit item Edit item
Provide Feedback