Mootz

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.

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
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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
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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
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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
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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
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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
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B. Di Ventura, H.D. Mootz
Biol. Chem., 2019, 400, 467-475
Switchable inteins for conditional protein splicing
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S. Palei, K.-S. Becher, C. Nienberg, J. Jose, H.D. Mootz
ChemBioChem2019, 20, 72-77
Bacterial cell surface display of semisynthetic cyclic peptides
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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
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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
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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.
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S. Palei, H.D. Mootz
Methods Mol. Biol. 2017, 1495, 77-92
Preparation of semisynthetic peptide macrocycles using split inteins.
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S. Palei, H. Mootz
ChemBioChem. 2016, 17(5), 378-382
Cyclic Peptides Made by Linking Synthetic and Genetically Encoded Fragments
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W. Fischle, H. Mootz, D. Schwarzer
Curr. Opin. Chem. Biol. 2015, 28, 131-140 (Review)
Synthetic histone code.
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W. Fischle, D. Schwarzer, H.D. Mootz
Nat. Chem. 2015, 7, 371-37
Chemical biology: Chromatin chemistry goes cellular
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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
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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
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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
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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
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