The main objective of our group is the study of genetic variability in drug (xenobiotic) metabolism and its role in susceptibility to therapeutic drugs and environmental agents. This variability is a major determining factor in the beneficial and adverse effects inchemical exposure.
Research concerns functional and mechanistic studies of drug metabolism enzyme variants to obtain insight in molecular mechanism of function of these enzymes, allowing the rationalization of their effect in exposure to agents and in specific disease states. We focus mainly on human cytochrome P450, one of the most important enzyme families in drug metabolism, using various in vitro models, and a pleiotropy of genetic and biochemical techniques.
Funding by Horizon Europe (HORIZON-HLTH-2022-STAYHLTH-02/101095679)
Collaboration with Prof. Javier Cubero (Complutense University of Madrid - coordinator)
Know more here.
The project regards an EU funded Cost Action which addresses a serious health problem in Europe namely drug induced liver injury or DILI. The objectives of the PRO-EURO-DILI-NET Cost Action are to create a unique, co-operative, interdisciplinary European-based DILI network of stakeholders to co-ordinate efforts in DILI, to facilitate bi-directional exchange of discovered knowledge and generated hypotheses among different disciplines, and to promote clinically impactful knowledge discovery and its translation into clinical practice.
This Action will:
The network will promote and coordinate a highly translational and innovative research program in Europe and beyond with the ultimate goal to pre-empt and prevent DILI, develop innovative therapeutic approaches that could improve clinical outcomes and enhance public awareness, while developing a forum for knowledge exchange and training of young European researches.Michel Kranendonk, the head of the Xenobiotic Metabolism lab, is member of the Management Committee of this Cost Action as well as member of the Workgroup 3.
Open-access publication:ADVANCED PRECLINICAL MODELS FOR EVALUATION OF DRUG INDUCED LIVER INJURY - CONSENSUS STATEMENT BY THE EUROPEAN DRUG-INDUCED LIVER INJURY NETWORK [PRO-EURO-DILI-NET] Checa et al. (2021) J Hepatol https://doi.org/10.1016/j.jhep.2021.06.021
NADPH cytochrome P450 oxidoreductase (CPR) is a diflavin protein which is the obligatory electron donor of CYP and non-CYP enzymes, which are involved in many physiological important processes. CPR, like all diflavin electron transfer (ET) enzymes, is a multidomain protein which undergoes large conformational changes in its ET function. CPR exists in a conformational equilibrium, between a closed (electron acceptor (NADPH) conformation and an open electron donation (interaction with redox partner) conformation. At the start of the project there were indications that both intrinsic (CPR determinants) and extrinsic (membrane environment and redox partners) control CPR’s open-closed dynamics, however several of these critical parameters were not analyzed.
The project had as goal to address these issues by:
The collaborative project between Lisbon and Toulouse (INSA) addressed these issues using both soluble purified (Partner 1, Toulouse) and membrane bound CPR forms (Partner 2 Lisbon).Several critical residues in a specific segment of the CPR protein, the hinge region could be identified in the open-closed dynamics of CPR, in which ionic strength seems to play a role. This extensive protein dynamics is responsible for the formation of an ensemble of open CPR conformers, by which CPR can interact with so many structurally different redox partners. Moreover, it was demonstrated that each redox partner (i.e. CYPs) interact with CPR via the use of specific biding-motifs of the interaction surface, the FMN domain, in an isoform specific manner. Additionally, we found that the substrate bound by CYP influences this interaction.
Team Members:Bernardo Brito Palma, Bruno Costa Gomes, Francisco Esteves, Diana Campelo
Esteves F, et al. Interaction Modes of Microsomal Cytochrome P450s with Its Reductase and the Role of Substrate Binding. Int. J. Mol. Sci. 2020 Sep 11;21(18):6669. doi: 10.3390/ijms21186669
Esteves F, etal. The Role of the FMN-Domain of Human Cytochrome P450 Oxidoreductase in Its Promiscuous Interactions with Structurally Diverse Redox Partners. Frontiers Pharmacol. 2020 Mar 18;11:299. doi: 10.3389/fphar.2020.00299
Campelo D, et al. Probing the Role of the Hinge Segment of Cytochrome P450 Oxidoreductase in the Interaction with Cytochrome P450. Int. J. Mol. Sci. 2018 Dec 6;19(12):3914. doi: 10.3390/ijms19123914
Quast RB. et al., Accurate Determination of Human CPR Conformational Equilibrium by smFRET Using Dual Orthogonal Noncanonical Amino Acid Labeling. Chembiochem. 2019 Mar 1;20(5):659-666. doi: 10.1002/cbic.201800607
Esteves F, et al. Human cytochrome P450 expression in bacteria: Whole-cell high-throughput activity assay for CYP1A2, 2A6 and 3A4. Biochem Pharmacol. 2018 Dec;158:134-140. doi: 10.1016/j.bcp.2018.10.006
Campelo D, et al. The Hinge Segment of Human NADPH-Cytochrome P450 Reductase in Conformational Switching: The Critical Role of Ionic Strength. Frontiers Pharmacol. 2017 Oct 30;8:755. doi: 10.3389/fphar.2017.00755
Esteves F, et al. The Central Role of Cytochrome P450 in Xenobiotic Metabolism—A Brief Review on a Fascinating Enzyme Family. J.Xenobiot. 2021, 11, 94–114. https://doi.org/10.3390/jox11030007
Deadline for contact: 10th March 2023For candidates residing in Portugal only.
Former Lab Members:
NOVA Medical SchoolCampo Mártires da Pátria, 1301169-056 LisboaPORTUGAL
GeralTel.: +351 218 803 000
Lista de Contactos
Elogios, Sugestões e Reclamações
Portal de Denúncias
Associação de Estudantes
Prémios e Distinções