Freund/Schwarzer

Establishing strategies for sortase-catalyzed multi-peptide assemblies to profile T-cell selectivity

The bacterial transpeptidase sortase A has become a versatile tool for protein chemistry by catalyzing chemoselective ligation reactions of peptides, proteins and synthetic probes. During catalysis sortase cleaves a so-called sorting motif consisting of the amino acid sequence LPxTG or LPxTA at the threonine residue under concomitant formation of a sortase-bound thioester. The enzyme is deacylated by ligating the thioester to the glycine residue of a second peptide that serves as nucleophile. While sortase A of S. aureus is widely used in biochemistry and chemical biology the assembly of multiple peptides is limited by the fact that each sorting motif and nucleophile is recognized by sortase, thereby leading to fragment shuffling and polymerization as a manifesting side reaction when fragment numbers increase. We plan to address these problems by establishing novel ligation sites to enable multi-fragment assemblies. We will use directed evolution methods to engineer sortase selectivity and activity. We plan to use these new tools for establishing tailor-made probes for immunology, in particular for the investigation of T-cell activation. 

Prof. Dr. Christian Freund
Freie Universität Berlin
Institut für Chemie und Biochemie
Thielallee 63

Tel. 030-838-51187
Fax. 030-838-56413

Email Prof. Freund

Prof. Dr. Dirk Schwarzer
Eberhard Karls Universität Tübingen

Tel.: +49 7071 29 - 73344
Fax: +49 7071 29 - 4815

Email Prof. Schwarzer

Publications within the SPP 1623 project

L. Schmohl, J. Bierlmeier, N. von Kügelgen, L. Kurz, P. Reis, F. Barthels, P. Mach, M. Schutkowski, C. Freund, D. Schwarzer
Bioorg Med. Chem. 2017, 25, 5002-5007
Identification of sortase substrates by specificity profiling
Link to the article


L. Schmohl, J. Bierlmeier, F. Gerth, C. Freund, D. Schwarzer
J Pept Sci. 2017, 23, 631-635
Engineering sortase A by screening a second-generation library using phage display
Link to the article


W. Fischle, D. Schwarzer
ACS Chemical Biology
2016, 11(3), 689-705 (Review)
Probing Chromatin-modifying enzymes with Chemical Tools.
Link to the article

A. Stützer, S. Liokatis, A. Kiesel, D. Schwarzer, R. Sprangers, J. Söding, P. Selenko, W. Fischle
Molecular Cell
2016, 61, 247-259
Modulations of DNA Contacts by Linker Histones and Post-translational Modifications Determine the Mobility and Modifiability of Nucleosomal H3 Tails.
Link to the article

A. Dose, J. Sindlinger, J. Bierlmeier, A. Bakirbas, K. Schulze-Osthoff, S. Einsele-Scholz, M. Hartl, F. Essmann, I. Finkemeier, D. Schwarzer
Angew. Chem. Int. Ed.
2016, 55, 1192-1195
Interrogating Substrate Selectivity and Composition of Endogenous Histone Deacetylase Complexes with Chemical Probes.
Link to the article

J. Seidel, C. Klockenbusch, D. Schwarzer
Chembiochem
2015, Ahead of print
Investigating deformylase and deacylase activity of mammalian and bacterial sirtuins.

W. Fischle, H. Mootz, D. Schwarzer
Curr. Opin. Chem. Biol.
2015, 28, 131-140 (Review)
Synthetic histone code.
Link to the article

O. Jost, A. Hanswillemenke, D. Schwarzer
Mol Biosyst.
2015, 11, 1820-1823
A miniaturized readout strategy for endogenous histone ceacetylase activity.
Link to the article

L. Schmohl, F.R. Wagner, M. Schümann, E. Krause, D. Schwarzer
Bioorg. Med. Chem.
2015, in press
Semisynthesis and initial characterization of sortase A mutants containing selenocysteine and homocysteine.
Link to the article

L. Schmohl, D. Schwarzer
Curr. Opin. Chem. Biol.
2014, 22, 122-128
Sortase-mediated ligations for the site-specific modification of proteins
Link to the article

K.A. Gelato, M. Tauber, M.S. Ong, S. Winter. K. Hiragami-Hamada, J. Sindlinger, A. Lemak, Y. Bultsma, S. Houliston, D. Schwarzer, N. Divecha, C.H. Arrowsmith, W. Fischle
Mol. Cell
2014, 54, 905-919
Accessibility of Different Histone H3-Binding Domains of UHRF1 Is Allosterically Regulated by Phosphatidylinositol 5-Phosphate.
Link to the article

L. Schmohl, D. Schwarzer
J. Pept. Sci.
2014, 20, 145-151
Chemo-enzymatic three-fragment assembly of semisynthetic proteins.
Link to the article