Targeted poly(ADP-ribosyl)ation of proteins

ADP-ribosylation and poly(ADP-ribosyl)ation are reversible posttranslational modifications of proteins catalyzed by ADP-ribosyltransferases (ARTs) like poly(ADP-ribose) polymerases (PARPs) using NAD+ as substrate. Poly(ADP-ribose) (PAR) is a linear or multibranched polyanion of variable size that can interact noncovalently with numerous proteins. PAR formation has been associated with DNA repair, maintenance of genomic stability, transcription, telomere regulation, cell division, energy metabolism, cell death, tumour suppression and ageing. However, our knowledge of how PAR and ADP-ribose modifications modulate the properties and regulate the function of their target proteins is sparse. 

For a better understanding of these modifications and their biological role we propose the synthesis of proteins with distinct sites of mono- and poly(ADP-ribosyl)ation. The generated functionalized proteins will be of major interest and the basis for comprehensive biochemical follow-up studies. By the incorporation of alkyne-modified unnatural amino acids via amber-stop-codon suppression, site-specific modification of proteins with orthogonally reacting functional groups is feasible. Subsequently, new ligation chemistry will be developed to offer a powerful tool to conjugate the unnatural amino acid site-specifically e.g. with azide-modified ADP-ribose and PAR. Such well-defined functionalized proteins will be used to investigate the biological role of mono- and poly(ADP-ribosyl)ation of the two exemplary histones H1 and H2.B, whose PARmodification has been revealed as a regulator of chromatin structure.

Prof. Dr. Alexander Bürkle
Universität Konstanz

Tel.: +49 7531 88 - 4035
Fax: +49 7531 88 - 4033

Email Prof. Bürkle

Prof. Dr. Andreas Marx
Universität Konstanz

Tel.: +49 7531 88 - 5139
Fax: +49 7531 88 - 5140

Email Prof. Marx

Publications within the SPP 1623 project

Y. Wang, D. Rösner, M. Grzywa, A. Marx
Angew. Chem. Int. Ed. 2014, 53, 8159-8162.
Chain terminating and clickable NAD+ analogues for labeling target proteins of ADP-ribosyltransferases
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A.-K. Späte, H. Bußkamp, A. Niederwieser, V. F. Schart, A. Marx, V. Wittmann
Bioconjug Chem.2014, 25, 147-154
Rapid Labeling of Metabolically Engineered Cell-Surface Glycoconjugates with a Carbamate-Linked Cyclopropene Reporter
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