Since biological tissues are unstable in an oxygen atmosphere, a great deal of effort is expended by organisms to metabolically limit or repair oxidative tissue damage. This volume of Methods in Enzymology and its companion Volume 233 present methods developed to investigate the roles of oxygen radicals and antioxidants in disease. It includes techniques and assays of the activity of biothiols in soluble systems, their interaction with lipids and proteins, and the various new thiol enzymes, such as thioltransferases, being elucidated. Antioxidant reactions and gene regulation are also included.

This volume of Methods in Enzymology and its companion Volume 238 include molecular, biological, and biochemical methods for the study of cell surface heterotrimeric G proteins (Volume 237) and their effectors (Volume 238). Methods unique to signal transducing G proteins and general techniques applied to the study of G protein systems are covered.

Hemoglobin has been involved in the most significant advances in our understanding of modern genetics and molecular biology. Now, hemoblobin is again central to a new area: development of artificial blood (blood substitute.) This volume of Methods in Enzymology and its companion, Volume 232, will be indispensable to anyone with a serious interest in this emerging field. They completely update and extend the information presented in Volume 76 published a decade ago.
Key Features
* Preparation, purification, and analysis of human hemoglobin and of various non-human hemoglobins
* Preparation of hemoglobin hybrids and of model heme compounds
* Preparation of chemically modified hemoglobins and of recombinant hemoglobins
* Analytical procedures presented for studying hemoglobin stability and degradation and for the formation of xenobiotic adducts of hemoglobin

The critically acclaimed laboratory standard, Methods in Enzymology, is one of the most highly respected publications in the field of biochemistry. Since 1955, each volume has been eagerly awaited, frequently consulted, and praised by researchers and reviewers alike. The series contains much material still relevant today - truly an essential publication for researchers in all fields of life sciences.

To understand the structures and dynamics of DNA and its interactions with ligands, a complete analysis of its confirmational flexibility is required. This volume and its companion, Volume 212 of Methods in Enzymology, collate in one source all the methods and approaches needed to achieve this goal.

The aim of this volume is to brief researchers of the importance of data analysis in enzymology, of the modern methods that have developed concomitantly with computer hardware, and of the need to validate their computer programs with real and synthetic data to ascertain that the results produced are what they expected.

This volume provides up-to-date information on the purification and characterization of the enzymes involved in phospholipid biosynthesis and on the mechanisms utilized by cells for the transfer of phospholipids. It also includes protocols for generating phospholipid biosynthetic mutants.

This book is aimed at biochemists, geneticists, molecular biologists, cell biologists, microbiologists, genetic engineers, analytical chemists, biophysicists, and industrial researchers working on protein products.;Of use to researchers who study transcriptional control, protein-DNA recognition, DNA replication, protein structure and function, and related topics, it aims to provide an up-to-date and critical description of the chemical, biochemical, biophysical, and genetic methods that are commonly used in the study of DNA binding proteins.

This volume provides up-to-date, state-of-the-art methods for determining the metal content of metalloenzymes, metalloproteins, and other metallobiochemicals. It emphasizes the need for control of contamination, techniques for handling and dealing with contamination-sensitive molecules, and the absolute necessity of quantitating metal content in systems inferred to contain metals by indirect means.

This volume collates in one source methodology for in vivo genetic engineering and for genetic analysis in a wide range of bacteria. Not only is "Escherichia coli" well covered, but so are other emerging bacterial systems.

This volume provides a compilation of recent methods for studying protein phosphorylation.

This volume on Phospholipases, the first of its kind, presents techniques for the assay, purification, and characterization of these enzymes, which are not only degradative enzymes of lipid metabolism but are also key enzymes in the signal transduction pathway.

The critically acclaimed laboratory standard, Methods in Enzymology, is one of the most highly respected publications in the field of biochemistry. Since 1955, each volume has been eagerly awaited, frequently consulted, and praised by researchers and reviewers alike. The series contains much material still relevant today - truly an essential publication for researchers in all fields of life sciences.

This volume is the first comprehensive compilation of methods for the assay and purification of enzymes involved in the utilization of reduced one-carbon compounds and higher hydrocarbons. In the last decade there has been an explosion of interest in the organisms that grow on one-carbon compounds and in those that grow on higher hydrocarbons. This is due to the commercial interest in the unique enzymes involved in these specialty metabolic pathways and to a growing understanding of the important role these organisms play in carbon cycling in nature. More recent interest in the use of these organisms and their enzymes for detoxification of hazardous waste has once again put the spotlight on these unique microorganisms. This volume will prove to be a valuable source for researchers in academia, government, and industry alike.

The critically acclaimed laboratory standard, Methods in Enzymology, is one of the most highly respected publications in the field of biochemistry. Since 1955, each volume has been eagerly awaited, frequently consulted, and praised by researchers and reviewers alike. The series contains much material still relevant today - truly an essential publication for researchers in all fields of life sciences.

The methods presented in this volume will enable the reader to design effective strategies for the expression of cloned genes and cDNAs and will prove useful in solving the majority of expression problems one is likely to encounter.

This volume addresses a variety of areas in which computers are used to manage and manipulate nucleic acid and protein sequence data. The manipulations include searching, aligning, and determining the significance of similarities, as well as the construction of phylogenetic trees that show the evolutionary history of related sequences. Ready-to-use methods for the "at-the-bench" scientist are presented.

Many methods have been developed or adapted for the study of RNA processing, which plays a key role in the expression of genetic information. Volumes 180 and 181 of Methods in Enzymology, RNA Processing, Parts A and B, represent the first comprehensive collation of protocols used for the preparation of substrates, for the preparation of extracts and enzymes, and for the analysis of products in RNA processing. At least half of the chapters present techniques that are directly applicable to other areas of nucleic acid biochemistry and cell biology.

This volume, as does Volume 177, provides a general background of modern NMR techniques, with a specific focus on NMR techniques that pertain to proteins and enzymology, and a "snapshot" of the current state of the art in NMR experimental techniques. These books enable the reader to understand a given technique, to evaluate its strengths and limitations, to decide which is the best approach, and, finally, to design an experiment using the chosen technique to solve a problem.

The critically acclaimed laboratory standard, Methods in Enzymology, is one of the most highly respected publications in the field of biochemistry. Since 1955, each volume has been eagerly awaited, frequently consulted, and praised by researchers and reviewers alike. The series contains much material still relevant today - truly an essential publication for researchers in all fields of life sciences.

This volume includes chapters on the newest techniques for measuring metabolites of branched-chain amino acids, many new enzyme assays such as the measurement of isozymes of acetohydroxacid synthase, and the purification of recently described enzymes such as isovaleryl-CoA dehydrogenase.

The critically acclaimed laboratory standard, Methods in Enzymology, is one of the most highly respected publications in the field of biochemistry. Since 1955, each volume has been eagerly awaited, frequently consulted, and praised by researchers and reviewers alike. The series contains much material still relevant today - truly an essential publication for researchers in all fields of life sciences.

The critically acclaimed laboratory standard, Methods in Enzymology, is one of the most highly respected publications in the field of biochemistry. Since 1955, each volume has been eagerly awaited, frequently consulted, and praised by researchers and reviewers alike. The series contains much material still relevant today - truly an essential publication for researchers in all fields of life sciences.

From the Preface
Although it has been suspected since ancient times that metals participate in biological processes, details of the manner in which they might exert a function have until recently remained unknown. It is now appreciated that metals play roles in catalysis, hormone action, gene and other regulatory functions, structural stabilization of macromolecules, muscle contraction, nerve conduction, and transport. "Metallobiochemistry" is a field which has grown and matured rapidly over the past decade. Metalloenzymes have been the central investigating targets. This volume provides both the emerging and practicing metallobiochemist with the necessary tools to answer what should be the number one and two questions: Is this enzyme a metalloenzyme? What metal is it?

Recombinant DNA methods are powerful, revolutionary techniques for at least two reasons.
First, they allow the isolation of single genes in large amounts from a pool of thousands or millions of genes. Second, the isolated genes or their regulatory regions can be modified at will and re-introduced into cells for expression at the RNA or protein levels. These attributes help to solve complex biological problems and to produce new and better products in the areas of health, agriculture and industry.
Both volume 154 and 155 include the descriptions of several useful new restriction enzymes and rapid methods for DNA sequence analysis, along with a number of other useful methods.