Ph.D. Student position in Biochemistry / Cell Biology / Endocrinology at University of Bern
PandeyLab: http://www.pandeylab.org University of Bern: http://www.unibe.ch
Graduate School of Cellular and Biomedical Sciences: http://www.gcb.unibe.ch
Group: https://www.dbmr.unibe.ch/research/research_programs/pandey_lab/index_eng.html
A fully funded Ph.D. Student position, funded by the Swiss National Science Foundation, is open in the laboratory of Amit Pandey at the University of Bern, starting April 2025 or later for research projects in Biochemistry / Cell Biology / Endocrinology. The student will work on Computational Redesign of human P450 oxidoreductase protein which combines cell and molecular biology, gene editing using CRISPR/Cas, and protein engineering experiments in Switzerland together with single molecule conformational studies with collaborators in Copenhagen, aiming to understand the mechanisms of electron transfer proteins and improve their stability and solubility for use in biotechnological applications.
Cytochrome P450 proteins metabolize drugs and xenobiotics in humans. A small number of P450 proteins are responsible for the metabolism of the majority of drugs on the market. Cytochrome P450 protein involved in drug metabolism requires another protein called P450 reductase for enzymatic activity. Previously, we have shown that P450 reductase in humans can have variations that alter the metabolic activities of several P450 proteins. Mutations in human NADPH cytochrome P450 oxidoreductase (POR) cause metabolic disorders. Recent work has set the foundation that POR does not act as a passive electron donor activating cytochrome P450s (CYPs), but rather operates as a central molecular hub selectively donating electrons and activating CYPs, controlling metabolic cascades. Previous research has focused on which POR mutations cause aberrant CYP functions and metabolic disorders. However, we have a very limited understanding of how POR mutations result in aberrant metabolism, which is key for controlling POR-mediated metabolic disorders.
The project will involve the use of the CRISPR/CAS system to edit the human P450 reductase sequence in cells to create models for the study of modified variants. This will allow the measurement of enzyme activities in an ideal cell environment. In a second approach, we will create recombinant forms of POR mutants by bacterial expression and purification, and assay enzyme kinetics and interaction with cytochrome P450 and other redox partners. The project would involve expressing recombinant P450 proteins in E. coli, purification of proteins by chromatography, and assay of enzyme activities using spectrometers. Further analysis of protein interactions will be done using ELISA and immuno-precipitation. Several protein and molecular biology techniques would be used to understand the molecular basis of POR conformational changes that affect metabolism due to variants in POR.
Job Requirements:
• Highly motivated and able to work both independently and in collaboration.
• Strong background in biochemistry / cell biology / molecular biology.
• Experience in human cells, recombinant protein expression, and enzyme assays is a plus.
• A willingness to work with a multidisciplinary team of international collaborators.
• Enthusiastic nature and good interpersonal and communication skills, with the ability to collaborate with other members of the team, including supervision of junior students.
• Engagement in lab activities including lab duties and presentation of research.
Formal requirements:
• Applicants should have a master’s degree in life sciences/biochemistry/cell biology that is acceptable for graduate studies at the University of Bern. For applicants who have not completed their master’s degree, please provide a confirmation from your supervisor that you will finish in time to join as a PhD student.
• Candidates must have good written and spoken communication skills in English, which is the working language of our laboratory.
• The applicant will have to be admitted and accepted by the Graduate School for Cellular and Biomedical Sciences at the University of Bern.
Offer: We offer a stimulating international working environment, state-of-the-art facilities, and integrative training, including molecular biology techniques, bioinformatic tools, and cell biological assays. University Children’s Hospital Bern is part of the medical faculty of the University of Bern. Salary will be according to the guidelines of the University of Bern and Swiss National Science Foundation and includes pension and social security benefits.
Application: Please send your application as a single pdf file, including a letter of motivation and curriculum vitae with reference letters and names of two referees to Prof. Dr. Amit V Pandey at: amit.pandey@unibe.ch
Related publications:
1. Flück, C. E.; Tajima, T.; Pandey, A. V.; Arlt, W.; Okuhara, K.; Verge, C. F.; Jabs, E. W.; Mendonca, B. B.; Fujieda, K.; Miller, W. L., Mutant P450 oxidoreductase causes disordered steroidogenesis with and without Antley-Bixler syndrome. Nat Genet 2004, 36, (3), 228-230.
2. Pandey, A. V.; Flück, C. E., NADPH P450 oxidoreductase: structure, function, and pathology of diseases. Pharmacol Ther 2013, 138, (2), 229-54.
3. Pandey, A. V.; Sproll, P., Pharmacogenomics of human P450 oxidoreductase. Front Pharmacol 2014, 5, 103.
4. Jensen, S. B.; Thodberg, S.; Parween, S.; Moses, M. E.; Hansen, C. C.; Thomsen, J.; Sletfjerding, M. B.; Knudsen, C.; Del Giudice, R.; Lund, P. M.; Castano, P. R.; Bustamante, Y. G.; Velazquez, M. N. R.; Jorgensen, F. S.; Pandey, A. V.; Laursen, T.; Moller, B. L.; Hatzakis, N. S., Biased cytochrome P450-mediated metabolism via small-molecule ligands binding P450 oxidoreductase. Nat Commun 2021, 12, (1), 2260.
5. Parween, S.; Fernandez-Cancio, M.; Benito-Sanz, S.; Camats, N.; Rojas Velazquez, M. N.; Lopez-Siguero, J. P.; Udhane, S. S.; Kagawa, N.; Fluck, C. E.; Audi, L.; Pandey, A. V., Molecular Basis of CYP19A1 Deficiency in a 46,XX Patient With R550W Mutation in POR: Expanding the POR Defect Phenotype. J Clin Endocrinol Metab 2020, 105, (4).
6. Parween, S.; DiNardo, G.; Baj, F.; Zhang, C.; Gilardi, G.; Pandey, A. V., Differential effects of variations in human P450 oxidoreductase on the aromatase activity of CYP19A1 polymorphisms R264C and R264H. J Steroid Biochem Mol Biol 2020, 196, 105507.
7. Parween, S.; Roucher-Boulez, F.; Flück, C. E.; Lienhardt-Roussie, A.; Mallet, D.; Morel, Y.; Pandey, A. V., P450 Oxidoreductase Deficiency: Loss of Activity Caused by Protein Instability From a Novel L374H Mutation. J Clin Endocrinol Metab 2016, 101, (12), 4789-4798.
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