G protein-dependent signalling by CXCR4 and CXCR7

Early stage researcher 7 (ESR7) project – Cristina Perpina
Supervision: Prof MJ Lohse, Prof C Hoffmann, Dr Isabella Maiellaro
Host: UWUE (DE) – University of Würzburg, Institute of Pharmacology and Toxicology and Rudolf Virchow Center

I- Project proposal:

Aim:

  1. Develop fluorescence resonance energy transfer (FRET)-based biosensors to monitor receptor and G protein activation.
  2. Identify the G protein subtypes activated by CXCR4 and CXCR7 mono-, homo-, and heteromers in response to different modulators.
  3. Develop FRET-based biosensors to monitor multiplex G protein effector signalling in real time.

Methodology:
This project aims to analyze signaling by chemokine receptors developing and using optical tools and methods. Biosensors shall be developed for both chemokine receptors (CXCR4 and CXCR7) and their G-proteins, which allow to monitor and image their activation by changes in fluorescence (FRET). These sensors will be expressed in cell lines (and possibly ultimately also in transgenic models) in order to image by FRET microscopy where and when the receptors become activated and how they signal. With the help of different new labeling strategies we will aim for multiplex signal analysis, i.e. to monitor multiple signaling pathways simultaneously.

Planned secondments: MPG (DE), Actelion (CH).

 

II- Requirement candidate:

Required diploma: MSc degree in life or natural sciences (e.g. biology, biochemistry, biophysics, pharmacy)

Required expertise: Molecular biology, cell culture, microscopy

Recommended expertise: Recombinant protein expression, molecular cloning, basic biophysics courses

Key publications:

  1. Vilardaga JP, Bünemann M, Krasel C, Castro M, Lohse MJ (2003) Measurement of the millisecond activation switch of G-protein-coupled receptors in living cells. Nature Biotechnology 21, 807-812
  2. Vilardaga JP, Nikolaev VO, Lorenz K, Ferrandon S, Zhuang Z, Lohse MJ (2008) Conformational cross-talk between alpha-2A-adrenergic and mu-opioid receptors controls cell signaling. Nature Chemical Biology 4, 126-131
  3. Hlavackova V, Zabel U, Frankova D, Bätz J, Hoffmann C, Prezeau L, Pin J-P, Blahos J, Lohse MJ (2012) Sequential inter- and intra-subunit rearrangements during asymmetric activation of dimeric metabotropic glutamate receptor 1. Science Sign. 5, ra59

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Contact details

Please contact us at:
info@oncornet.eu

ONCORNET Coordinator
VU University Amsterdam
The Netherlands