Computational and Experimental Studies of ADP-Ribosylation

Methods Mol Biol. 2017:1608:475-513. doi: 10.1007/978-1-4939-6993-7_29.

Abstract

The macrodomains are a multifunctional protein family that function as receptors and enzymes acting on poly(ADP-ribose), ADP-ribosylated proteins, and other metabolites of nicotinamide adenine dinucleotide (NAD+). Several new functions for macrodomains, such as nucleic acid binding and protein-protein interaction, have recently been identified in this family. Here, we discuss methods for the identification of new macrodomains in viruses and the prediction of their function. This is followed by the expression and purification of these proteins following overexpression in bacterial cells and confirmation of folding and function using biophysical methods.

Keywords: Bioinformatics; Circular dichroism; Coronavirus; Docking; Expression; Macrodomain; NMR spectroscopy; Purification; STD-NMR.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, Non-U.S. Gov't
  • Research Support, N.I.H., Extramural

MeSH terms

  • ADP-Ribosylation
  • Circular Dichroism
  • Computational Biology / methods*
  • Coronavirus / genetics
  • Coronavirus / metabolism
  • Magnetic Resonance Spectroscopy
  • Protein Binding / genetics
  • Protein Binding / physiology
  • Protein Processing, Post-Translational