Terfenadine metabolism of human cytochrome P450 2J2 containing genetic variations (G312R, P351L and P115L)
| Autor: | Yejin Lee, Young-Ran Lim, Hyoung-Goo Park, Vitchan Kim, Myung-A. Cho, Donghak Kim, Dabin Jeong |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Histamine H1 Antagonists Non-Sedating Pharmaceutical Science medicine.disease_cause Cytochrome P-450 CYP2J2 Polymorphism Single Nucleotide Protein Structure Secondary Hydroxylation 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Cytochrome P-450 Enzyme System medicine Humans Pharmacology (medical) Enzyme kinetics Escherichia coli Heme Pharmacology chemistry.chemical_classification biology Active site Cytochrome P450 Genetic Variation Metabolism 030104 developmental biology Enzyme chemistry Biochemistry 030220 oncology & carcinogenesis biology.protein Terfenadine |
| Zdroj: | Drug metabolism and pharmacokinetics. 33(1) |
| ISSN: | 1880-0920 |
| Popis: | The human cytochrome P450 2J2 is involved in several metabolic reactions, including the oxidation of important therapeutics and epoxidation of endogenous arachidonic acid. At least ten genetic variations of P450 2J2 have been identified, but their effects on enzymatic activity have not been clearly characterized. Here, we evaluated the functional effects of three genetic variations of P450 2J2 (G312R, P351L, and P115L). Recombinant enzymes of wild-type and three variant P450 2J2 were heterologously expressed in Escherichia coli and purified. P450 expression levels in the wild-type and two variants (P351L and P115L) were 142–231 nmol per liter culture, while the G312R variant showed no holoenzyme peak in the CO-binding spectra. Substrate binding titrations to terfenadine showed that the wild-type and two variants displayed Kd values of 0.90–2.2 μM, indicating tight substrate binding affinities. Steady-state kinetic analysis for t-butyl methyl hydroxylation of terfenadine indicated that two variant enzymes had similar kcat and Km values to wild-type P450 2J2. The locations of mutations in three-dimensional structural models indicated that the G312R is located in the I-helix region near the formal active site in P450 2J2 and its amino acid change affected the structural stability of the P450 heme environment. |
| Databáze: | OpenAIRE |
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