Protein Sequencing Protocols

Beschreibung

Bryan Smith draws together in Protein Sequencing Protocols a timely collection of current and state-of-the-art techniques for protein and peptide preparation and sequencing-all described by hands-on masters of the procedures. These well-tested methods convey valuable lessons and insights into using automated sequencers and/or mass spectrometers, preparing proteins and peptides on a microgram and submicrogram scale, mapping peptides, analyzing amino acid by pre- and post-column derivatization methods, and sequencing N- and C-terminal peptides by classical and newly developed techniques. They also deal with such troublesome problems as identifying sites of modification and handling very small samples.

Protein Sequencing Protocols offers self-contained, step-by-step recipes that ensure easily reproducible results for those already doing sequencing, and for those who are still learning the methods, as well as for those already in one area but wishing to learn associated techniques. Its multiplicity of proven protocols will guide the challenged experimenter to successful results.
If the development of techniques for the labeling of a polypeptide- terminus and for repetitive N-terminal sequencing mark the beginning of the science of protein structure determination, then the field has just about reached its half century. In more recent times, recombinant DNA techniques have pro vided powerful means by which to obtain long protein sequences (by theoreti cal translation from nucleic acid sequences), but rather than replacing the direct, chemical, protein sequencing approach, they have instead added further impe tus to the drive towards a better understanding of posttranslational processing and modification events, as well as identification of novel proteins. Recent years have also seen the advent of "biopharmaceuticals" (i. e. , pharmaceutical products that are proteins, such as monoclonal antibodies), and this has meant that protein sequencing has found an important new application as a quality control tool. Over the decades of protein sequencing many new techniques have been introduced, with the basic aim of generating more information from less material. Some techniques have come and gone, but others have been with us for many years. Edman chemistry is perhaps the best example of the latter class, with its basic principles still being applied today in the newest protein sequencer design. Methods for cleaving and modifying peptides are also long standing, having been adapted over the years to suit progressively smaller amounts of sample.

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Bryan John Smith

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"This methodological guide can be recommended to all who are concerned with protein characterization or peptide mapping."-Reviewed by R. Moravec, Biologia Plantarum
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