Protein Sequencing and Identification Using Tandem Mass Spectrometry

By addebook • Jun 22nd, 2008 • Category: Biology •

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Protein Sequencing and Identification Using Tandem Mass Spectrometry

Protein Sequencing and Identification Using Tandem Mass Spectrometry
By Michael Kinter, Nicholas E. Sherman

Publisher: Wiley-Interscience
Number Of Pages: 320
Publication Date: 2000-09-18
ISBN-10 / ASIN: 0471322490
ISBN-13 / EAN: 9780471322498
Binding: Hardcover

How to design, execute, and interpret experiments for protein sequencing using mass spectrometry

The rapid expansion of searchable protein and DNA databases in recent years has triggered an explosive growth in the application of mass spectrometry to protein sequencing. This timely and authoritative book provides professionals and scientists in biotechnology research with complete coverage of procedures for analyzing protein sequences by mass spectrometry, including step-by-step guidelines for sample preparation, analysis, and data interpretation.

Michael Kinter and Nicholas Sherman present their own high-quality, laboratory-tested protocols for the analysis of a wide variety of samples, demonstrating how to carry out specific experiments and obtain fast, reliable results with a 99uccess rate. Readers will get sufficient experimental detail to apply in their own laboratories, learn about the proper selection and operation of instruments, and gain essential insight into the fundamental principles of mass spectrometry and protein sequencing. Coverage includes:

Peptide fragmentation and interpretation of product ion spectra
Basic polyacrylamide gel electrophoresis
Preparation of protein digests for sequencing experiments
Mass spectrometric analysis using capillary liquid chromatography
Techniques for protein identification by database searches
Characterization of modified peptides using tandem mass spectrometry
And much more

Summary: Protein Sequencing and Identification Using Tandem Mass Spectrometry
Rating: 5

Ao ler esse livro pude perceber que existe variadas técnicas que auxiliam na interpretação dos espectros de massas. A obra descreve os métodos de ionização e a formação dos íons moleculares e quasi-moleculares em cada um desses métodos. O livro tras a interpretação da distribuição de isótopos nas amostras, a identificação dos “parent ions”, “base piks” e a distribuição das cargas durante a interpretação dos espectros. A interpretação de sequências de aminoácidos durante a análise de peptídios é rica em dicas, tabelas e exercícios resolvidos. De forma clara é possível entender a lógica por tras da formação das séries de íons A, B, C, X, Y e Z tão importantes durante a identificação da estrutura primária dos peptídeos. Se você pretende trabalhar com a identidficação de compostos de origem protéica utilizando espectrometria de massas, esse deverá ser seu livro de cabeceira!

Summary: Broad, thorough introduction
Rating: 5

I’m not a bench chemist, but I needed a quick survey of how mass spectroscopy is used in handling proteins and other big biomolecules. This book was it.

Although brief, it is thorough and well-organized. The first two chapters are mostly an introduction. Chapter 1 states the problem being solved. The next chapter briefly introduces older technologies, including chemical techiques and 60s-80s mass spec technique. The next two chapters summarize modern mass spec hardware, then start to show how proteins behave in the environment inside the instrument. That gives the fundamentals of protein sequencing: how the molecules break down, and how the fragments help recreate the molecule. The authors go through a few examples in detail, starting from a mass spectrogram and moving forward to sequence. I was especially impressed by the examples that fail. Mass spec analysis is not a magic wand for producing sequences, it is a deductive process, and can not complete an analysis when clues are missing or ambiguous.

The next three chapters are not about mass spec directly. Instead, they discuss how samples are prepared for analysis. This includes the clearest, most informative description of gel electrophoresis that I’ve seen, along with features of gel chemistry that do or do not interfere with mass spec measurements. This includes a discussion of protein digests, enzymatically produced fragments, and their place in analysis. I would have liked a little more discussion about combining information from digests produced by different enzymes, but no book can cover everything.

The last three chapters extend the discussion of analysis, working upwards from fragments to complete protein sequences. The three chapters respectively address three topics: using standard internet databases for recognizing fragments of known proteins, using combinations of strategies to analyze novel proteins, and using mass spec to identify post-translational modifications. That last one suffers from brevity; perhaps it was only meant to define a problem that deserves a whole book of its own.

Despite its throughness, the authors resist the urge for boggling detail. They present detail up to the point needed for understanding the mechanism and meaning of their topics, then stop. Lots of other writing would benefit from that kind of restraint.

I came away from this book well-informed, and ready to address specific topics in greater detail. That was exactly what I wanted. I recommend this book very highly.

//wiredweird

Summary: “Protein Sequencing ……” a must to read
Rating: 5

Michael Kinter has presented the topic in a scientific yet enjoyable format. I found the information to be extremely interesting and beneficial in my laboratory. Lets face it folks this isn’t the easiest nor most interesting topic to write about. Job well done DR. Kinter. I only have one criticism, there should have been more photos and illustrations. Get your copies quick this will no doubt be on the NewYork times best seller list before long.

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