| Chapter title |
Biochemical and Biophysical Assays of PAR-WWE Domain Interactions and Production of iso-ADPr for PAR-Binding Analysis
|
|---|---|
| Chapter number | 5 |
| Book title |
ADP-ribosylation and NAD+ Utilizing Enzymes
|
| Published in |
Methods in molecular biology, August 2018
|
| DOI | 10.1007/978-1-4939-8588-3_5 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-8587-6, 978-1-4939-8588-3
|
| Authors |
Zhizhi Wang, Wenqing Xu, Wang, Zhizhi, Xu, Wenqing |
| Abstract |
The poly(ADP-ribose) polymerase (PARP) family of proteins utilize NAD+ as the substrate to modify protein acceptors with either mono(ADP-ribose) (MAR) or poly(ADP-ribose) (PAR). MAR and PAR have been shown to regulate distinct cellular processes. Iso-ADP-ribose (iso-ADPr) is the smallest internal PAR structural unit containing the characteristic ribose-ribose glycosidic bond formed during poly(ADP-ribosyl)ation. The WWE domain of RNF146 specifically recognizes the iso-ADPr moiety in PAR but does not interact with MAR. This provides a way to distinguish PAR from MAR modification and to isolate PARylated proteins. Iso-ADPr can be used to detect the PAR-specific binding properties of interested proteins. Here we describe the detailed method to generate and purify iso-ADPr and its use in PAR-binding analysis through isothermal titration calorimetry (ITC) analysis. |
Mendeley readers
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 5 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Student > Master | 2 | 40% |
| Professor > Associate Professor | 1 | 20% |
| Student > Ph. D. Student | 1 | 20% |
| Unknown | 1 | 20% |
| Readers by discipline | Count | As % |
|---|---|---|
| Biochemistry, Genetics and Molecular Biology | 2 | 40% |
| Agricultural and Biological Sciences | 1 | 20% |
| Chemistry | 1 | 20% |
| Unknown | 1 | 20% |