Ando, Y., Elkayam, E., McPherson, R. L., Dasovich, M., Cheng, S. J., Voorneveld, J., Filippov, D. V., Ong, S. E., Joshua-Tor, L., Leung, A. K. L. (January 2019) ELTA: Enzymatic Labeling of Terminal ADP-Ribose. Mol Cell, 73 (4). 845-856 e5. ISSN 1097-2765
Abstract
ADP-ribosylation refers to the addition of one or more ADP-ribose groups onto proteins. The attached ADP-ribose monomers or polymers, commonly known as poly(ADP-ribose) (PAR), modulate the activities of the modified substrates or their binding affinities to other proteins. However, progress in this area is hindered by a lack of tools to investigate this protein modification. Here, we describe a new method named ELTA (enzymatic labeling of terminal ADP-ribose) for labeling free or protein-conjugated ADP-ribose monomers and polymers at their 2'-OH termini using the enzyme OAS1 and dATP. When coupled with various dATP analogs (e.g., radioactive, fluorescent, affinity tags), ELTA can be used to explore PAR biology with techniques routinely used to investigate DNA or RNA function. We demonstrate that ELTA enables the biophysical measurements of protein binding to PAR of a defined length, detection of PAR length from proteins and cells, and enrichment of sub-femtomole amounts of ADP-ribosylated peptides from cell lysates.
| Item Type: | Paper |
|---|---|
| Subjects: | bioinformatics bioinformatics > genomics and proteomics > genetics & nucleic acid processing bioinformatics > genomics and proteomics Investigative techniques and equipment bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification |
| CSHL Authors: | |
| Communities: | CSHL labs > Joshua-Tor lab CSHL Cancer Center Program > Gene Regulation and Inheritance Program |
| Depositing User: | Matthew Dunn |
| Date: | 25 January 2019 |
| Date Deposited: | 05 Feb 2019 21:40 |
| Last Modified: | 01 Feb 2024 20:08 |
| PMCID: | PMC6629254 |
| Related URLs: | |
| URI: | https://repository.cshl.edu/id/eprint/37686 |
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