Practical Protein Chemistry A Handbook

Protein chemistry is a wide field embracing the analysis, synthesis, sequencing and separation of peptides, proteins and individual amino acids. Practical protein chemistry is a survey of proven techniques designed primarily to be a laboratory handbook for research workers involved with proteins.

The Protein Protocols Handbook, Second Edition aims to provide a cross-section of analytical techniques commonly used for proteins and peptides, thus providing a benchtop manual and guide for those who are new to the protein chemistry laboratory and for those more established workers who wish to use a technique for the first time. All chapters are written in the same format as that used in the Methods in Molecular BiologyTM series. Each chapter opens with a description of the basic theory behind the method being described. The Materials section lists all the chemicals, reagents, buffers, and other materials necessary for carrying out the protocol. Since the principal goal of the book is to provide experimentalists with a full account of the practical steps necessary for carrying out each protocol successfully, the Methods section contains detailed st- by-step descriptions of every protocol that should result in the successful execution of each method. The Notes section complements the Methods material by indicating how best to deal with any problem or difficulty that may arise when using a given technique, and how to go about making the widest variety of modifications or alterations to the protocol. Since the first edition of this book was published in 1996 there have, of course, been significant developments in the field of protein chemistry.

As protein science continues to become an increasingly important aspect of academic and commercial sciences and technology, the need has arisen for a ready source of laboratory protocols for the analysis and evaluation of these biological polymers. Methods for Protein Analysis presents the methods most relevant to the generalist bench scientist working with proteins. A concise yet thorough summary, it covers laboratory methods that can be reasonably performed in a standard protein laboratory, without specialized equipment or expertise. Taking a how to approach, this book examines the techniques used to answer common protein analytical questions and describes methods useful in daily laboratory work. Methods for Protein Analysis is the ideal reference for protein laboratories in academic, government and industrial settings. It is an essential benchtop manual for first-year graduate students beginning their laboratory experience as well as for chemists, biochemists, and molecular biologists in the pharmaceutical, biotechnological, food and specialty chemical industries, and for analysts concerned with the purity and structural integrity of protein. Featuring illustrations and a convenient spiral binding, this guide offers a glossary of common abbreviations and a list of suppliers for protein science.

Milestones in the techniques and methodology of polypeptide structure determination include the determination of the sequence of insulin by Sanger in 1951 (I) and the introduction of the repeti tive degradation of proteins with phenylisothiocyanate by Edman in 1959 (2). The automation of Edman chemistry (3) played a major role in the determination of polypeptide structures. Important modifications of Edman chemistry include the solid-phase approach by Laursen in 1971 (4) and the use of modified Edman reagents such as 4-N, N-dimethylaminoazobenzene-4'-isothiocy- ate (DABITC) for manual sequencing by Chang et al. (5) in 1976. A second major breakthrough in the analysis of polypeptides was automated amino acid analysis described by Spackman et al. in 1958 (6). However, during the period from 1975 to 1980, it became increasingly clear that the amount of material required for struc tural analysis was more than could be easily isolated for the vast majority of proteins. The field was criticized for its lack of sensitive techniques for the analysis of growth factors, immune modulators, membrane receptors, and peptide hormones. In addition, very little had been done to modernize and improve the original instruments introduced in the mid-1960s. The first indications of improved instrumentation for Edman chemistry came from Wittmann-Liebold's laboratory (7), followed by the introduction of a "micro" sequencer by Hunkapiller and Hood in 1978 (8). The movement toward improved instrumentation culminated in the "gas"--Phase sequencer of Hewick et al. (9) in 1981

New textbooks at alllevels of chemistry appear with great regularity. Some fields like basic biochemistry, organic reaction mechanisms, and chemical thermodynamics are weil represented by many excellent texts, and new or revised editions are published sufficiently often to keep up with progress in research. However, some areas of chemistry, especially many of those taught at the graduate level, suffer from a reallack ofup-to-date textbooks. The most serious needs occur in fields that are rapidly changing. Textbooks in these subjects usually have to be written by scientists actually involved in the research which is advancing the field. lt is not often easy to persuade such individuals to settime aside to help spread the knowledge they have accumu lated. Our goal, in this series, is to pinpoint areas of chemistry where recent progress has outpaced what is covered in any available textbooks, and then seek out and persuade experts in these fields to produce relatively concise but instructive introductions to their fields. These should serve the needs of one semester or one quarter graduate courses in chemistry and biochemistry. In some cases the availability of texts in active research areas should help stimulate the creation of new courses. NewYork CHARLES R. CANTOR Preface to the Second Edition The original plan for the first edition of this book was to title it Enzyme Purification: Princip/es and Practice.

In recent years, interest in proteins has surged. This resurgence has been driven by the expansion of the post-genomic era when structural genomics and proteomics require new techniques in protein chemistry and new applications of older techniques. Protein chemistry methods are used by nearly every discipline of biomedical research. Many techniques