Identifying and Validating Tankyrase Binders and Substrates: A Candidate Approach.
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Date
2017-01-01Author
Pollock, K
Ranes, M
Collins, I
Guettler, S
Type
Journal Article
Metadata
Show full item recordAbstract
The poly(ADP-ribose)polymerase (PARP) enzyme tankyrase (TNKS/ARTD5, TNKS2/ARTD6) uses its ankyrin repeat clusters (ARCs) to recognize degenerate peptide motifs in a wide range of proteins, thereby recruiting such proteins and their complexes for scaffolding and/or poly(ADP-ribosyl)ation. Here, we provide guidance for predicting putative tankyrase-binding motifs, based on the previously delineated peptide sequence rules and existing structural information. We present a general method for the expression and purification of tankyrase ARCs from Escherichia coli and outline a fluorescence polarization assay to quantitatively assess direct ARC-TBM peptide interactions. We provide a basic protocol for evaluating binding and poly(ADP-ribosyl)ation of full-length candidate interacting proteins by full-length tankyrase in mammalian cells.
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Subject
Telomere
Animals
Humans
Tankyrases
Binding Sites
Protein Binding
Poly ADP Ribosylation
Research team
Medicinal Chemistry 2
Structural Biology of Cell Signalling
Language
eng
Date accepted
2017-01-01
License start date
2017-01
Citation
Methods in molecular biology (Clifton, N.J.), 2017, 1608 pp. 445 - 473
Publisher
Springer New York