New Proluciferin Substrates for Human CYP4 Family Enzymes
- PDF / 2,220,179 Bytes
- 20 Pages / 439.37 x 666.142 pts Page_size
- 75 Downloads / 195 Views
New Proluciferin Substrates for Human CYP4 Family Enzymes Jingyao Liu 1 & David Machalz 2 & Gerhard Wolber 2 & Erik J. Sorensen 1,3 & Matthias Bureik 1 Received: 3 March 2020 / Accepted: 16 July 2020/ # Springer Science+Business Media, LLC, part of Springer Nature 2020
Abstract
We report the synthesis of seven new proluciferins for convenient activity determination of enzymes belonging to the cytochrome P450 (CYP) 4 family. Biotransformation of these probe substrates was monitored using each of the twelve human CYP4 family members, and eight were found to act at least on one of them. For all substrates, activity of CYP4Z1 was always highest, while that of CYP4F8 was always second highest. Site of metabolism (SOM) predictions involving SMARTCyp and docking experiments helped to rationalize the observed activity trends linked to substrate accessibility and reactivity. We further report the first homology model of CYP4F8 including suggested substrate recognition residues in a catalytically competent conformation accessed by replica exchange solute tempering (REST) simulations. Keywords Cytochrome P450 . CYP4Z1 . Docking . Fission yeast . Homo sapiens . Pharmacology . Proluciferin . Site of metabolism prediction
Electronic supplementary material The online version of this article (https://doi.org/10.1007/s12010-02003388-6) contains supplementary material, which is available to authorized users.
* Erik J. Sorensen [email protected] * Matthias Bureik [email protected]
1
School of Pharmaceutical Science and Technology, Health Sciences Platform, Tianjin University, Tianjin 300072, China
2
Pharmaceutical and Medicinal Chemistry (Computer-Aided Drug Design), Institute of Pharmacy, Freie Universität Berlin, 14195 Berlin, Germany
3
Department of Chemistry, Princeton University, Princeton, NJ 08544, USA
Applied Biochemistry and Biotechnology
Introduction The large superfamily of cytochrome P450 enzymes (CYPs or P450s) can be found in many different organisms, with numbers per genome ranging from two in the fission yeast Schizosaccharomyces pombe to several hundreds in many plants [1]. These hemoproteins are best known for catalyzing hydroxylation reactions and show a huge substrate spectrum [2]. In humans, there are 57 functional CYP enzymes [3], which are involved in the metabolization of xenobiotics (such as active pharmaceutical ingredients, food-derived chemicals, environmental toxins, and carcinogens) and in the biosynthesis, metabolism, and degradation of endogenous compounds (like fatty acids and steroids) [4]. Several of these CYPs (for instance, aromatase) are drug targets, and some CYP inhibitors are already on the market, while others are being tested in clinical trials [5, 6]. Human CYP4Z1 was originally identified due to its overexpression in breast cancer tissue [7]. Shortly afterwards, the presence of positive CYP4Z1 immunoreactivity in primary ovarian cancer was found to be associated with poor prognosis [8]. After recombinant expression in the fission yeast Schizosaccharomyces pombe, we could dem
Data Loading...
