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ADP-ribosylation and NAD+ Utilizing Enzymes

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Cover of 'ADP-ribosylation and NAD+ Utilizing Enzymes'

Table of Contents

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    Book Overview
  2. Altmetric Badge
    Chapter 1 Vitamin B3 in Health and Disease: Toward the Second Century of Discovery
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    Chapter 2 Monitoring Poly(ADP-Ribosyl)ation in Response to DNA Damage in Live Cells Using Fluorescently Tagged Macrodomains
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    Chapter 3 In Vitro Techniques for ADP-Ribosylated Substrate Identification
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    Chapter 4 Assessment of Intracellular Auto-Modification Levels of ARTD10 Using Mono-ADP-Ribose-Specific Macrodomains 2 and 3 of Murine Artd8
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    Chapter 5 Biochemical and Biophysical Assays of PAR-WWE Domain Interactions and Production of iso-ADPr for PAR-Binding Analysis
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    Chapter 6 Assays for NAD+-Dependent Reactions and NAD+ Metabolites
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    Chapter 7 Generating Protein-Linked and Protein-Free Mono-, Oligo-, and Poly(ADP-Ribose) In Vitro
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    Chapter 8 Methods to Study TCDD-Inducible Poly-ADP-Ribose Polymerase (TIPARP) Mono-ADP-Ribosyltransferase Activity
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    Chapter 9 Dictyostelium as a Model to Assess Site-Specific ADP-Ribosylation Events
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    Chapter 10 Mono-ADP-Ribosylation Catalyzed by Arginine-Specific ADP-Ribosyltransferases
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    Chapter 11 Monitoring Expression and Enzyme Activity of Ecto-ARTCs
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    Chapter 12 ADP-Ribosyl-Acceptor Hydrolase Activities Catalyzed by the ARH Family of Proteins
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    Chapter 13 Mono-ADP-Ribosylhydrolase Assays
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    Chapter 14 Hydrolysis of ADP-Ribosylation by Macrodomains
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    Chapter 15 HPLC-Based Enzyme Assays for Sirtuins
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    Chapter 16 Small-Molecule Screening Assay for Mono-ADP-Ribosyltransferases
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    Chapter 17 A Simple, Sensitive, and Generalizable Plate Assay for Screening PARP Inhibitors
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    Chapter 18 Nonlocalized Searching of HCD Data for Fast and Sensitive Identification of ADP-Ribosylated Peptides
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    Chapter 19 Quantitative Determination of MAR Hydrolase Residue Specificity In Vitro by Tandem Mass Spectrometry
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    Chapter 20 Detection of ADP-Ribosylating Bacterial Toxins
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    Chapter 21 Preparation of Recombinant Alphaviruses for Functional Studies of ADP-Ribosylation
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    Chapter 22 Monitoring the Sensitivity of T Cell Populations Towards NAD+ Released During Cell Preparation
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    Chapter 23 Identifying Target RNAs of PARPs
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    Chapter 24 ADPr-Peptide Synthesis
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    Chapter 25 Identifying Genomic Sites of ADP-Ribosylation Mediated by Specific Nuclear PARP Enzymes Using Click-ChIP
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    Chapter 26 Methods for Using a Genetically Encoded Fluorescent Biosensor to Monitor Nuclear NAD +
Attention for Chapter 21: Preparation of Recombinant Alphaviruses for Functional Studies of ADP-Ribosylation
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Chapter title
Preparation of Recombinant Alphaviruses for Functional Studies of ADP-Ribosylation
Chapter number 21
Book title
ADP-ribosylation and NAD+ Utilizing Enzymes
Published in
Methods in molecular biology, August 2018
DOI 10.1007/978-1-4939-8588-3_21
Pubmed ID
Book ISBNs
978-1-4939-8587-6, 978-1-4939-8588-3

Rachy Abraham, Robert Lyle McPherson, Easwaran Sreekumar, Anthony K. L. Leung, Diane E. Griffin


Recently we characterized the mono(ADP-ribosyl) hydrolase (MAR hydrolase) activity of the macrodomain of nonstructural protein 3 (nsP3MD) of chikungunya virus. Using recombinant viruses with targeted mutations in the macrodomain, we demonstrated that hydrolase function is important for viral replication in cultured neuronal cells and for neurovirulence in mice. Here, we describe the general cell culture and animal model infection protocols for alphaviruses and the technical details for biochemical characterization of the MAR hydrolase activity of nsP3MD mutants and the preparation of recombinant viruses incorporating those mutations through site-directed mutagenesis of an infectious cDNA virus clone.

Mendeley readers

The data shown below were compiled from readership statistics for 4 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
Unknown 4 100%

Demographic breakdown

Readers by professional status Count As %
Student > Postgraduate 1 25%
Researcher 1 25%
Unknown 2 50%
Readers by discipline Count As %
Agricultural and Biological Sciences 2 50%
Unknown 2 50%

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