Universität zu Köln

Bonfiglio, Juan Jose ORCID: 0000-0001-7767-0799, Leidecker, Orsolya, Dauben, Helen, Longarini, Edoardo Jose, Colby, Thomas, San Segundo-Acosta, Pablo, Perez, Kathryn A. and Matic, Ivan ORCID: 0000-0003-0170-7991 (2020). An HPF1/PARP1-Based Chemical Biology Strategy for Exploring ADP-Ribosylation. Cell, 183 (4). S. 1086 - 1126. CAMBRIDGE: CELL PRESS. ISSN 1097-4172

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Abstract

Strategies for installing authentic ADP-ribosylation (ADPr) at desired positions are fundamental for creating the tools needed to explore this elusive post-translational modification (PTM) in essential cellular processes. Here, we describe a phospho-guided chemoenzymatic approach based on the Ser-ADPr writer complex for rapid, scalable preparation of a panel of pure, precisely modified peptides. Integrating this methodology with phage display technology, we have developed site-specific as well as broad-specificity antibodies to monoADPr. These recombinant antibodies have been selected and characterized using multiple ADP-ribosylated peptides and tested by immunoblotting and immunofluorescence for their ability to detect physiological ADPr events. Mono-ADPr proteomics and poly-to-mono comparisons at the modification site level have revealed the prevalence of mono-ADPr upon DNA damage and illustrated its dependence on PARG and ARH3. These and future tools created on our versatile chemical biology-recombinant antibody platform have broad potential to elucidate ADPr signaling pathways in health and disease.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Bonfiglio, Juan Jose UNSPECIFIED orcid.org/0000-0001-7767-0799 UNSPECIFIED Leidecker, Orsolya UNSPECIFIED UNSPECIFIED UNSPECIFIED Dauben, Helen UNSPECIFIED UNSPECIFIED UNSPECIFIED Longarini, Edoardo Jose UNSPECIFIED UNSPECIFIED UNSPECIFIED Colby, Thomas UNSPECIFIED UNSPECIFIED UNSPECIFIED San Segundo-Acosta, Pablo UNSPECIFIED UNSPECIFIED UNSPECIFIED Perez, Kathryn A. UNSPECIFIED UNSPECIFIED UNSPECIFIED Matic, Ivan UNSPECIFIED orcid.org/0000-0003-0170-7991 UNSPECIFIED
URN:	urn:nbn:de:hbz:38-311543
DOI:	10.1016/j.cell.2020.09.055
Journal or Publication Title:	Cell
Volume:	183
Number:	4
Page Range:	S. 1086 - 1126
Date:	2020
Publisher:	CELL PRESS
Place of Publication:	CAMBRIDGE
ISSN:	1097-4172
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language COMPUTATIONAL PLATFORM; RIBOSE; ANTIBODIES; SERINE; IDENTIFICATION; EXPRESSION; PHOSPHORYLATION; PURIFICATION; GENERATION; MECHANISM Multiple languages Biochemistry & Molecular Biology; Cell Biology Multiple languages
URI:	http://kups.ub.uni-koeln.de/id/eprint/31154