Chapter title |
Identifying and Validating Tankyrase Binders and Substrates: A Candidate Approach
|
---|---|
Chapter number | 28 |
Book title |
Poly(ADP-Ribose) Polymerase
|
Published in |
Methods in molecular biology, July 2017
|
DOI | 10.1007/978-1-4939-6993-7_28 |
Pubmed ID | |
Book ISBNs |
978-1-4939-6992-0, 978-1-4939-6993-7
|
Authors |
Katie Pollock, Michael Ranes, Ian Collins, Sebastian Guettler |
Abstract |
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. |
Mendeley readers
Geographical breakdown
Country | Count | As % |
---|---|---|
Unknown | 18 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Student > Ph. D. Student | 3 | 17% |
Researcher | 3 | 17% |
Other | 1 | 6% |
Student > Bachelor | 1 | 6% |
Professor | 1 | 6% |
Other | 5 | 28% |
Unknown | 4 | 22% |
Readers by discipline | Count | As % |
---|---|---|
Biochemistry, Genetics and Molecular Biology | 7 | 39% |
Agricultural and Biological Sciences | 2 | 11% |
Chemistry | 2 | 11% |
Social Sciences | 1 | 6% |
Unspecified | 1 | 6% |
Other | 0 | 0% |
Unknown | 5 | 28% |