Chemoselective DNA conjugation of proteins to study multienzyme complexes

This project concerns the development of a generic platform to generate multfunctional protein constructs useful for interrogation of biological systems. The platform technology is based on the combination of chemoselective protein ligation methods and structural DNA nanotechnology. In particular, we will establish the chemoenzymatic ligation of recombinant proteins derived from multimodular Polyketide Synthases (PKS) with DNA or PNA oligonucleotides, which are derivatized with a short peptide tag, serving as the ligation substrate of bacterial transpeptidase Sortase. 

The resulting DNA-/PNA-tagged PKS modules will be assembled by Watson-Crick base pairing in homogeneous solution and, in particular, on the surface of two-dimensional nucleic acid scaffolds generated by the DNA origami technique. In a highly modular approach, supramolecular multienzyme complexes will be built, in which the spatial configuration of the PKS domains is controlled by the DNA scaffold with nanometer precision.

On the one hand, this project will contribute to the detailed understanding of the spatial organization, conformational flexibility and other fundamental principles governing multienzyme complexes. On the other hand, the platform developed here will enable ready access to numerous spatially well-defined multifunctional protein complexes for mimicking, manipulation and analysis of natural occuring supramolecular assemblies involved in gene regulation, signal transduction, division, or other cellular function and traits.

Prof. Dr. Christof Niemeyer
Karlsruher Institut für Technologie (KIT)

Tel.: +49 721 608 23000
Fax: +49 721 608 25546

Email Prof. Niemeyer

Prof. Dr. Frank Schulz
Organic Chemistry I
Faculty of Chemistry and Biochemistry
Ruhr-University Bochum

Tel.: +49 234-32-27034

Email Prof. Schulz

Publications within the SPP 1623 project

C. Timm, C.M. Niemeyer
Angew Chem Int Ed2015, 54, 6745-6750
Assembly and Purification of Enzyme-Functionalized DNA Origami Structures
Link to the article

R. Meyer, A. Faesen, K. Vogel, S. Jeganathan, A. Musacchio, C.M. Niemeyer
Small2015, 11(22), 2669-2674
DNA-Directed Assembly of Capture Tools for Constitutional Studies of Large Protein Complexes.
Link to the article

S. Klopries, K. Koopmans, E. Sanchez-Garcia, F. Schulz
ChemBioChem2014, 15(4), 495-497
Biosynthesis with Flourine
Link to the article