Nuclease Methods and Protocols (Methods in Molecular Biology, Vol. 160) - Hardcover

Hardcover

ISBN 10: 0896036790 ISBN 13: 9780896036796

Publisher: Humana, 2001

Synopsis

Nucleases, enzymes that restructure or degrade nucleic acid polymers, are vital to the control of every area of metabolism. They range from “housekeeping” enzymes with broad substrate ranges to extremely specific tools (1). Many types of nucleases are used in lab protocols, and their commercial and clinical uses are expanding. The purpose of Nuclease Methods and Protocols is to introduce the reader to some we- characterized protein nucleases, and the methods used to determine their activity, structure, interaction with other molecules, and physiological role. Each chapter begins with a mini-review on a specific nuclease or a nuclease-related theme. Although many chapters cover several topics, they were arbitrarily divided into five parts: Part I, “Characterizing Nuclease Activity,” includes protocols and assays to determine general (processive, distributive) or specific mechanisms. Methods to assay nuclease products, identify cloned nucleases, and determine their physiological role are also included here. Part II, “Inhibitors and Activators of Nucleases,” summarizes assays for measuring the effects of other proteins and small molecules. Many of these inhibitors have clinical relevance. Part III, “Relating Nuclease Structure and Function,” provides an overview of methods to determine or model the 3-D structure of nucleases and their complexes with substrates and inhibitors. A 3-D structure can greatly aid the rational design of nucleases and inhibitors for specific purposes. Part IV, “Nucleases in the Clinic,” summarizes assays and protocols suitable for use with t- sues and for nuclease based therapeutics.

About this title

About the Author

The editor of Nuclease Methods and Protocols, Catherine H. Schein, PhD, obtained degrees in chemistry (U. Pennsylvania), biochemical engineering (MIT) and technical microbiology (ETH Zurich). After pioneering work in producing recombinant proteins in bacteria, including the first large (100,000 l) scale production of human interferon for clinical studies, she started investigating the role of nucleases in cytokine mechanisms. Her work on protein solubility eventually lead her to computational and structural biology. She currently coordinates projects between medical research groups in the Sealy Center for Structural Biology at one of the largest medical research centers in the United States, the University of Texas Medical Branch at Galveston

The authors of the individual chapters are from academic and industrial research groups from four different continents.

From the Back Cover

Nucleases, the enzymes that cleave and shape nucleic acids, are key regulators of cellular metabolism. Angiogenesis and the unfolded protein response are just a few instances of important intra- and extracellular pathways regulated by nucleases. In Nuclease Methods and Protocols, researchers at the forefront of academic research and pharmaceutical drug development from four continents summarize their recent results. Their projects involve nucleases in many different areas of medicine and biology, ranging from protein structure and folding, to DNA repair, to developing cures for insidious diseases such as cystic fibrosis, AIDS, and cancer. Each chapter contains a minireview of a specific nuclease or nuclease-related theme, a discussion of why and when to use the assays described, and often, an explanation of how the assay was developed. The authors describe their most valuable methods for determining the mode of action, structure, interaction with other molecules, and physiological role of nucleases. Detailed step-by-step instructions and notes on avoiding pitfalls help ensure readily reproducible results.
Comprehensive and highly practical, Nuclease Methods and Protocols offers
both novice and experienced researchers a deeper understanding of the importance of nucleases in cell metabolism, growth control mechanisms, and, increasingly, in
diagnostics and therapeutics.

From the Inside Flap

Nucleases, the enzymes that cleave and shape nucleic acids, are key regulators of cellular metabolism. Angiogenesis and the unfolded protein response are just a few instances of important intra- and extra-cellular pathways regulated by nucleases. Nuclease Methods and Protocols provides an introduction to this extensive family of enzymes and well-polished methodologies for studing and exploiting their activities. Groups at the forefront of academic research and pharmaceutical drug development, from four continents, summarize their recent results. Their projects involve nucleases in many different areas of medicine and biology, ranging from protein structure and folding to DNA repair to developing cures for insidious diseases such as cystic fibrosis, AIDS and cancer. Many novel diagnostic methods and therapies are described in the section of the book devoted to clinical uses of nucleases.

Each chapter contains a minireview of a specific nuclease or nuclease-related theme, a discussion of why and when to use the assays described, and often, an explanation of how the assay was developed. The authors describe their most valuable methods for determining the mode of action, structure, interaction with other molecules, and physiological role of nucleases. Separate sections of the book describe assays for nucleases and for characterizing their interaction with inhibitors and activators, determining structure, and preparing modified protein nucleases for clinical and diagnostic purposes. Detailed step-by-step instructions and notes on avoiding pitfalls help ensure readily reproducible results.

Comprehensive and highly practical, Nuclease Methods and Protocols offers both novice and experienced researchers a deeper understanding of the importance of nucelases in cell metabolism, growth control mechanisms, and, increasingly, in therapeutics.

Edition details

• Publisher: Humana
• Publication date: 2001
• Language: English
• ISBN 10: 0896036790
• ISBN 13: 9780896036796
• Binding: Hardcover
• Number of pages: 542
• Editor: Schein Catherine H.