ADP-ribosylation and NAD+ Utilizing Enzymes: Methods and Protocols

by Paul Chang

Overview

This volume focuses on mono-ADP-ribosylation and enzymes that use NAD+ including Sirtuins, PARPs, and bacterial and eukaryotic ADP-ribosyltransferases. The chapters in this book are organized into eight parts, and offer detailed descriptions of key prools used to study topics such as in vitro techniques for ADP-ribosylation substrate identification; biochemical and biophysical assays of PAR-WWE domain interactions; monitoring expression and enzyme activity of ecto-ARTCs; HPLC-based enzymes assays for Sirtuins; and identifying target RNAs of PARPs. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory prools, and tips on troubleshooting and avoiding known pitfalls.

Cutting-edge and thorough, ADP-ribosylation and NAD+ Utilizing Enzymes: Methods and Prools is a valuable resource for anyone interested in this developing and expanding field.

Product Details

ISBN-13:	9781493985876
Publisher:	Springer New York
Publication date:	08/11/2018
Series:	Methods in Molecular Biology , #1813
Edition description:	1st ed. 2018
Pages:	417
Product dimensions:	7.01(w) x 10.00(h) x (d)

Table of Contents

Vitamin B3 in Health and Disease: Toward the Second Century of Discovery.- Monitoring Poly(ADP-Ribosyl)ation in Response to DNA Damage in Live Cells Using Fluorescently-Tagged Macrodomains.- In Vitro Techniques for ADP-Ribosylated Substrate Identification.- Assessment of Intracellular Auto-Modification Levels of ARTD10 using Mono-ADP-Ribose-Specific Macrodomains 2 and 3 of Murine Artd8.- Biochemical and Biophysical Assays of PAR-WWE Domain Interactions and Production of iso-ADPr for PAR Binding Analysis.- Assays for NAD-Dependent Reactions and NAD Metabolites.- Generating Protein-Linked and Protein-Free Mono-, Oligo-, and Poly(ADP-Ribose) In Vitro.- Methods to Study TCDD-Inducible Poly-ADP-Ribose Polymerase (TIPARP) Mono-ADP-Ribosyltransferase Activity.- Dictyostelium as a Model to Assess Site-Specific ADP-Ribosylation Events.- Mono-ADP-Ribosylation Catalyzed by Arginine-Specific ADP-Ribosyltransferases.- Monitoring Expression andEnzyme Activity of Ecto-ARTCs.- ADP-Ribosyl-Acceptor Hydrolase Activities Catalyzed by the ARH Family of Proteins.- Mono-ADP-Ribosylhydrolase Assays.- Hydrolysis of ADP-Ribosylation by Macrodomains.- HPLC-Based Enzyme Assays for Sirtuins.- Small Molecule Screening Assay for Mono-ADP-Ribosyltransferases.- A Simple, Sensitive, and Generalizable Plate Assay for Screening PARP Inhibitors.- Non-Localized Searching of HCD Data for Fast and Sensitive Identification of ADP-Ribosylated Peptides.- Quantitative Determination of MAR Hydrolase Residue Specificity In Vitro by Tandem Mass Spectrometry.- Detection of ADP-Ribosylating Bacterial Toxins.- Preparation of Recombinant Alphaviruses for Functional Studies of ADP-Ribosylation.- Monitoring the Sensitivity of T Cell Populations towards NAD+ Released during Cell Preparation.- Identifying Target RNAs of PARPs.- ADPr-Peptide Synthesis.- Identifying Genomic Sites of ADP-Ribosylation Mediated by Specific Nuclear PARP Enzymes Using Click-ChIP.- Methods for Using a Genetically-Encoded Fluorescent Biosensor to Monitor Nuclear NAD+.