Chemoselective Staudinger-induced Michael-additions to antibodies to analyze protein homeostasis in C. elegans

In this proposal a novel metal-free chemoselective conjugation strategy will be applied to modify antibodies for the analysis of protein-protein interactions of key relevance in the maintenance of protein homeostasis. In this joint effort the expertise of the Hackenberger laboratory in the engineering of bioorthogonal reactions and the Kirstein laboratory in protein homeostasis will be combined to identify novel chaperone complexes involved in protein disaggregation and their protein substrates in C. elegans. In addition, we aim to employ antibody drug conjugates to target modifiers of amyloid formation to Abeta formed fibrils in vivo. With the development of a Staudinger-induced Michael-addition for the chemoselective conjugation of cysteine residues in antibodies a new concept for the modification of proteins will be introduced.

This novel technique enables the modular and metal-free coupling of two complex functional building blocks without the need of intermediate protecting group manipulations. Additionally, a phosphonamidate moiety is introduced by this method, which allows the direct incorporation of a light- or enzymatically cleavable bond in protein conjugates. This new synthetic tool will be applied to obtain functional antibodies for the identification of unknown protein substrates as well as interaction partners of specific chaperones and will furthermore be used in the design of light-cleavable antibody drug conjugates to target amyloid fibrils in living animal models for Alzheimer's disease that express Abeta1-42 peptides.

Prof. Dr. Christian Hackenberger
Humboldt Universität zu Berlin
Leibniz-Institut für Molekulare Pharmakologie (FMP)

Tel.: +49 30 94793 - 181
Fax: +49 30 94793 - 188

Email Prof. Hackenberger

Dr. Janine Kirstein
Leibniz-Institut für Molekulare Pharmakologie (FMP)

Tel.: +49 30 94793 - 250
Email Dr. Kirstein

Publications within the SPP 1623 project

C.E. Stieger, L. Franz, F. Körlin, C.P.R. Hackenberger
Angew. Chem. Int. Ed. 2021, 60(28), 15359-15364
Diethynyl Phosphinates for Cysteine-Selective Protein Labeling and Disulfide Rebridging
Link to the article
Angew. Chem. 2021, 133(28), 15487-15492
Diethinylphosphinate für die Cystein-selektive Proteinmarkierung und Disulfid-Verbrückung
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Y. Park*, A.L. Baumann*, H. Moon, S. Byrne, M.-A. Kasper, S. Hwang, H. Sun, M.-H. Baik, C.P.R. Hackenberger
Chem. Sci. 2021, 12, 8141-8148
The mechanism behind enhanced reactivity of unsaturated phosphorus(v) electrophiles towards thiols
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P. Ochtrop, C.P.R. Hackenberger
Curr. Op. in Chem. Biology 2020, 58, 28-36
Recent advances of thiol-selective bioconjugation reactions (review)
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A.L. Baumann, S. Schwagerus, K. Broi, K. Kemnitz-Hassanin, C.E. Stieger, N. Trieloff, P. Schmieder, C.P.R. Hackenberger
JACS 2020, 142(20), 9544-9552
Chemically Induced Vinylphosphonothiolate Electrophiles for Thiol-Thiol Bioconjugations
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highlighted in: Synfacts 2020, 16(07), 0851, DOI: 10.1055/s-0040-1707878
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C. Gallrein, M. Iburg, T. Michelberger, A. Kocak, D. Puchkov, F. Liu, S.M. Ayala Mariscal, T. Nayak, G.S. Kaminski Schierle, J. Kirstein
Progress in Neurobiology 2020, 198, 101907
Novel Amyloid-beta pathology C. elegans model reveals distinct neurons as seeds of pathogenicity
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M. Iburg, D. Puchkov, I.U.R. Brugada, L. Bergemann, U. Rieprecht, J. Kirstein
J Biol Chem2020, 295(10), 3064-3079
The non-canonical small heat shock protein HSP-17 from C. elegans is a selective protein aggregase
Link to the article

A.L. Baumann, C.P.R. Hackenberger
Curr. Opin. Chem. Biol. 2019, 52, 39-46
Tag and release: strategies for the intracellular cleavage of protein conjugates (Review)
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D.R. Feleciano, K. Juenemann, M. Iburg, I. Bras, C.I. Holmberg, J. Kirstein
Frontiers in Aging Neuroscience 2019, Jan 29;11:9
Crosstalk between chaperone-mediated protein disaggregation and proteolytic pathways in aging and disease
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A. Baumann, C.P.R. Hackenberger
CHIMIA 2018, 72(11), 802-808
Modern Ligations Methods to Access Natural and Modified Proteins
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A. Scior, K. Arnsburg, M. Iburg, K. Juenemann, M.L. Pigazzini, B. Mlody, D. Puchkov, A. Ast, A. Buntru, J. Priller, E.E. Wanker, A. Prigione, J. Kirstein
The EMBO Journal 2018, 37, 282-299.
Complete suppression of Htt fibrilization and disaggregation of Htt fibrils by a chaperone complex.
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J. Kirstein, K. Arnsburg, A. Scior, A. Szlachcic, D.L. Guilbride, R. I. Morimoto, B. Bukau, N. B. Nillegoda
Aging Cell 2017, 16(6), 1414-1424
In vivo properties of the disaggregase function of J-proteins and Hsc70 in Caenorhabditis elegans stress and aging
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M.C. Lechler, E.D. Crawford, N. Groh, K. Widmaier, R. Jung, J. Kirstein, J.C. Ttrinidad. A.L. Burlingame. D.C. David
Cell Rep. 2017, 18(2), 454-467
Reduced Insulin/IGF-1 Signaling Restores the Dynamic Properties of Key Stress Granule Proteins during Aging.
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