Conformational signature of CXCR4/ACKR3 activation in β-arrestin-1 and -2

Early stage researcher 8 (ESR8) project
Supervision: Prof. Dr. Carsten Hoffmann, Dr. Julia Drube
Host: Universitätsklinikum Jena

I – Project proposal:

Our current knowledge is very limitted with respect to GPCR/arrestin interaction and the role of individual GRKs in this process. Most of the GPCR-family members are regulated by only four of the seven members of the GRK-family (namely GRK 2,-3,-5, and -6) and two of the four arrestin proteins that exist in human physiology, namely β-arrestin 1 and β-arrestin 2. For this system of GPCR regulation to be conceivable, β-arrestins require the ability to adopt to a multitude of different receptor activation and phosphorylation states, supposedly facilitated by a high degree of conformational plasticity within β-arrestin. 


  1. Characterization of the ligand dependent conformational signature of CXCR4/ACKR3 in β-arrestin-1 and -2 using FRET and BRET sensors for conformational changes in β-arrestin-1/2.
  2. Characterization of phosphorylation dependent conformational signature of CXCR4/ACKR3 in β-arrestin-1/-2 using site-directed mutagenesis and GRK specific knock-down (CRISPR/Cas9) in HEK293 cells. 
  3. The conformational signatures obtained will further be evaluated for differential signaling and/or localization of the respective chemokine receptor.

We will use a combination of site-directed mutagenesis of identified phosphorylation sites, pharmacological tools (GRK2 or GRK5 inhibition), and knock-down approaches (siRNA or CRISPR/Cas9 for GRKs using HEK293 cells) to study the individual effect of phosphorylation sites in coupling to β-arrestin.

Furthermore, FRET and BRET-based sensors will be developed and employed to study concentration dependent conformational changes of arrestin in real time and living cells. Confocal microscopy (including high-resolution) will be used to study the individual effects of phosphorylation sites in receptor localization. 

Planned secondments:  

Universidad Autónoma de Madrid (Prof. Federico Mayor)

Vrije Universiteit Amsterdam (Prof. Dr. Martine Smit)


II – Requirement candidate: 

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

Required expertise: biochemistry, molecular biology

Recommended expertise: Knowledge of basic cell culture, microscopy and imaging techniques (fluorescence of bioluminescence resonance energy transfer). Basic knowledge of ImageJ, Prism, Origin would be a plus. Knowledge of siRNA or CRISR/Cas9 would be a bonus.

Key publications:

  1. S. Nuber, U. Zabel, K. Lorenz, A. Nuber, G. Milligan, A.B. Tobin, M.J. Lohse, C. Hoffmann (2016) β-Arrestin biosensors reveal a rapid, receptor-dependent activation/deactivation cycle. Nature Mar 31;531(7596):661-4; doi: 10.1038/nature17198. Epub 2016 Mar 23.

  2. R.S. Haider, A. Goodbole, C. Hoffmann (2019) To sence or not to sence – New insights from GPCR and arrestin biosensors. Current opinion in Cell Biology, 57: 16-24 doi: 10.1016/

  3. J. Heuninck, C. Perpiñá Viciano, A. Işbilir, B. Caspar, D. Capoferri, S.J. Briddon, T. Durroux, S.J. Hill, M.J. Lohse, G. Milligan, J.-P. Pin, and C. Hoffmann (2019) Context-dependent signalling of CXC chemokine receptor 4 (CXCR4) and atypical chemokine receptor 3 (ACKR3) MolPharm, in press, doi: 10.1124/mol.118.115477

For more information:
Prof. Dr. Carsten Hoffmann –



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Great way to finish the week off with some #FCS measurements of CXCR4-expressing HEK cells at @UoNLifeSci 🔬 #FluorescenceFriday

Hello everyone!
I am Noemi Karsai and I’m the next in our #ESR introduction series. I’m #ESR11 and I’m originally from Hungary, currently doing my PhD at @UoNLifeSci @COMPARE_UoBUoN, UK.

The transatlantic ECI GPCR symposium #ECIGPCR is about to start, we are ready!! Thanks to the organizers for their excellent job gathering almost 500 people across the globe! @cyclic_Andreas @NicoleAPerry1 @BenderSci @DesislavaNeshe1

My project will be focused on the role of #CXCR4/ACKR3/GRK2 -governed networks in cancer cell migration and metastasis.

Hello 🙂
I am here to continue the series in which all #ESRs are presenting themselves. I am Viviana Marolda and I am in my first year of PhD. I am #ESR13, originally from Italy, and currently, I am working as a PhD student in @CBMSO_CSIC_UAM, at Universidad Autonoma de Madrid.

Hey twitter!
I am @DehanComez. I am taking over the Oncornet account for a while to start a series of all #ESRs writing about themselves and their projects. I am #ESR5, originally from Turkey and right now I am working as a PhD student in @COMPARE_UoBUoN , @UniofNottingham , UK.

New review on #gpcr structural dynamics out in COSB. Great teamwork with the @JanaSelent group. Thanks to @Lundbeckfonden and @novonordiskfond

ONCORNET2.0 is the successor to #ONCORNET. You can see some of the work of ESRs from the first ONCORNET in this special issue of @MolPharmJournal from 2019, with reviews on #CXCR4 and #ACKR3 structure and function:

Hi everyone – we’re on Twitter! ONCORNET2.0 is a #MarieCurie ITN of 16 ESRs across Europe studying #chemokine #GPCRs #CXCR4 and #ACKR3 in cancer. Our projects cover molecular dynamics, medchem, #pharmacology through to translational work. Follow us for updates from our ESRs!

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Contact details

Please contact us at:

ONCORNET Coordinator
VU University Amsterdam
The Netherlands