Mootz

New approaches for the site-selective chemical modification of therapeutic proteins under mild conditions

The site-specific chemical modification of protein therapeutics, for example antibodies, protein hormones, and cytokines, is of great importance for applications in cancer and disease treatment, diagnostics, and imaging. Chemical modifications can endow the protein therapeutics with new and tailored properties, for example longer circulation time in the blood-stream, targeted cell-killing (immunoconjugates), and reporting their localization (e.g. by fluorophores or tracers). 

In the proposed project new methodologies will be developed that overcome several limitations of the currently most widely used classical bioconjugation approaches. The new conjugation strategies are based on protein trans-splicing catalyzed by split inteins and the combination with selected chemo- and regioselective reactions. They will enable site-specific single and double modification of proteins obtained from different expression host organisms. The target proteins will not be exposed to harsh chemical reaction conditions of any kind, but will rather be modified at pH 7 and even in the absence of reducing agents. While the current project will focus on the modification of antibodies, the methodologies developed here will also be applicable to in principle any other protein and thus significantly advance the repertoire of protein chemistry.

Prof. Dr. Henning Mootz
Westfälische Wilhelms-Universität Münster

Tel.: +49 251 83 - 33005

Email Prof. Mootz

Publications within the SPP 1623 project

S. Palei, H.D. Mootz
Chem. Comm.2021, 57, 4194-4197
Semisynthetic head-to-tail cyclized peptides obtained by combining protein trans-splicing and intramolecular expressed protein ligation
Link to the article

S. Hoffmann, M.E. Terhorst, R.K. Singh, D. Kümmel, S. Pietrokovski*, H.D. Mootz*
ChemBioChem 2021, 22, 364-373
Biochemical and structural characterization of an unusual and naturally split class 3 intein
Link to the article

M. Bhagawati, S. Hoffmann, K.S. Höffgen, J. Piehler, K.B. Busch, H.D. Mootz
Angew. Chem. Int. Ed. 2020, 59, 21007-21015
In cellulo protein semi-synthesis from endogenous and exogenous fragments using the ultra-fast split Gp41-1 intein
Link to the article

D. Aparicio Pelaz, Z. Yerkesh, S. Kirchgäßner, H. Mahler, V. Kharchenko, D. Azhibek, M. Jaremko, H.D. Mootz, Ł. Jaremko, D. Schwarzer, W. Fischle
Chem. Sci. 2020, 11, 9218-9225
Examining histone modification crosstalk using immobilized libraries established from ligation-ready nucleosomes
Link to the article

M. Bhagawati, T.M.E. Terhorst, F. Füsser, S. Hoffmann, T. Pasch, S. Pietrokovski*, H.D. Mootz*
Proc. Natl. Acad. Sci. USA2019, 116, 22164-22172
A mesophilic cysteine-less split intein for protein trans-splicing applications under oxidizing conditions
Link to the article

M. Reille-Seroussi, S.V. Mayer, W. Dörner, K. Lang and H.D.D. Mootz*
Chem. Comm.2019, 55, 4793-4796
Expanding the Genetic Code with a Lysine Derivative Bearing an Enzymatically Removable Phenylacetyl Group
Link to the article

B. Di Ventura, H.D. Mootz
Biol. Chem., 2019, 400, 467-475
Switchable inteins for conditional protein splicing
Link to the article

S. Palei, K.-S. Becher, C. Nienberg, J. Jose, H.D. Mootz
ChemBioChem2019, 20, 72-77
Bacterial cell surface display of semisynthetic cyclic peptides
Link to the article

J.C.J. Matern, K. Friedel, J. Binschik, K.-S. Becher, Z. Yilmaz, H.D. Mootz
J. Am. Chem. Soc.2018, 140, 11267-11275
Altered coordination of individual catalytic steps in different and evolved inteins reveals kinetic plasticity of the protein splicing pathway
Link to the article

T. Pirzer, K.S. Becher, M. Rieker, T. Meckel, H.D. Mootz, H. Kolmar
ACS Chem. Biol.2018, 13, 2058-2066
Generation of potent anti-HER1/2 immunotoxins by protein ligation using split inteins
Link to the article

A.-L. Bachmann, H.D. Mootz
J. Pep. Sci 2017, 23, 624-630.
N-terminal chemical protein labeling using the naturally split GOS-TerL intein.
Link to the article

S. Palei, H.D. Mootz
Methods Mol. Biol. 2017, 1495, 77-92
Preparation of semisynthetic peptide macrocycles using split inteins.
Link to the article

S. Palei, H. Mootz
ChemBioChem. 2016, 17(5), 378-382
Cyclic Peptides Made by Linking Synthetic and Genetically Encoded Fragments
Link to the article

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

W. Fischle, D. Schwarzer, H.D. Mootz
Nat. Chem. 2015, 7, 371-37
Chemical biology: Chromatin chemistry goes cellular
Link to the article

A.-L. Bachmann, J.C. Matern, V. Schütz, H.D. Mootz
Methods Mol. Biol. 2015, 1266, 145-159
Chemical-tag labeling of proteins using fully recombinant split inteins
Link to the article

J.C. Matern, A.-L. Bachmann, I.V. Thiel, G. Volkmann, A. Wasmuth, J. Binschik, H.D. Mootz
Methods Mol. Biol. 2015, 1266, 129-143
Ligation of Synthetic Peptides to Proteins Using Semisynthetic Protein trans-Splicing
Link to the article

V. Schütz, H.D. Mootz
Angew. Chem. Int. Ed. 2014, 126 (16), 4197-4201
Click-tag and amine-tag: New chemical tag approaches for efficient protein labeling in vitro and on live cells using the naturally split Npu DnaE intein
Link to the article

I.V. Thiel, G. Volkmann, S. Pietrokovski, H.D. Mootz
Angew. Chem. Int. Ed.2014, 53, 1306-1310
An atypical naturally split intein engineered for highly efficient protein labeling
Link to the article