Kubick/Schiller

Reconstitution of tyrosine kinase signaling in cell-free systems: Synthetic membrane protein dimerization and lipid modification

The Main Goals of the project are:

  • Reconstitution of EGFR depended signaling pathways in cell-free systems
  • In vitro generation and chemical modification of constitutively active EGF-Receptor (via genetically encoded unnatural amino acids utilized to dimerize the receptor permanently) and its downstream effector RhoA (via the introduction of lipid moieties facilitated by unnatural amino acids)
  •  Reconstitution of integral membrane proteins and membrane associated proteins in lipid bilayers by combination of chemical methods and naturally occurring posttranslational modifications in cell-free systems (via genetically encoded unnatural amino acids and novel heterobisfunctional linker chemistries)
  • Development of novel heterobisfunctional linker chemistries for protein dimerization- Implementation of novel tRNA/tRNA synthetase pairs for the incorporation of unnatural amino acids into proteins in various in vitro translation systems
  • Monitoring functionality of in vitro synthesized and constitutive active membrane proteins through detection of phosphorylation (Antibody based Assays) and Protein-Protein interaction analysis (mass spectrometry Assays)- Identification of novel inhibitory components with regard to the EGF-receptor’s downstream signaling pathway.

Dr. Stefan Kubick
Fraunhofer Institute for Cell Therapy and Immunology (IZI)
Branch Bioanalytics and Bioprocesses (IZI-BB), Potsdam
Department of Cell-free Bioproduction

Tel.: +49 331 58187 - 306
Fax: +49 331 58187 - 399

Email Dr. Kubick

Dr. Stefan Schiller
Freiburg Institute for Advanced Studies
Albert-Ludwigs-Universität Freiburg

Tel.: +49 761 203 97405
Fax: +49 761 203 97451

Email Dr. Schiller

Publications within the SPP 1623 project

K. Heller, P. Ochtrop, M.F. Albers, F.B. Zauner, A. Itzen, C. Hedberg
Angew. Chem. Int. Ed. 2015, accepted
Covalent protein labeling by enzymatic phosphocholination
 

D. Wiegandt, S. Vieweg, F. Hofmann, D. Koch, F. Li, Y. Wu, A. Itzen, M.P. Müller, R.S. Goody
Nature Communications 2015, accepted
Locking GTPases covalently in their functional states

C. Hedberg, A. Itzen
ACS Chem. Biol. 2015, 10, 12-21
Molecular perspectives on protein adenylylation.
Link to the article

M.P. Müller, M.F. Albers, A. Itzen, C. Hedberg
ChemBioChem2014, 15(1), 19-26
Exploring Adenylylation and Phosphocholinaton as Post-Translational Modifications
Link to the article

M.F. Albers, C. Hedberg
J. Org. Chem. 2013, 78(6), 2715-2719
Amino Acid Building Blocks for Fmoc Solid-Phase Synthesis of Peptides Phosphocholinated at Serine, Threonine, and Tyrosine
Link to the article