ELTA: Enzymatic Labeling of Terminal ADP-Ribose

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

URL: https://www.ncbi.nlm.nih.gov/pubmed/30712989
DOI: 10.1016/j.molcel.2018.12.022

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: Investigative techniques and equipment
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification
CSHL Authors:
Communities: CSHL labs > Joshua-Tor lab
Depositing User: Matthew Dunn
Date: 25 January 2019
Date Deposited: 05 Feb 2019 21:40
Last Modified: 25 Feb 2019 19:45
Related URLs:
URI: http://repository.cshl.edu/id/eprint/37686

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