Mass Spectrometry In Structural Biology And Biophysics

The definitive guide to mass spectrometry techniques in biology and biophysics The use of mass spectrometry (MS) to study the architecture and dynamics of proteins is increasingly common within the biophysical community, and Mass Spectrometry in Structural Biology and Biophysics: Architecture, Dynamics, and Interaction of Biomolecules, Second Edition provides readers with detailed, systematic coverage of the current state of the art. Offering an unrivalled overview of modern MS-based armamentarium that can be used to solve the most challenging problems in biophysics, structural biology, and biopharmaceuticals, the book is a practical guide to understanding the role of MS techniques in biophysical research. Designed to meet the needs of both academic and industrial researchers, it makes mass spectrometry accessible to professionals in a range of fields, including biopharmaceuticals. This new edition has been significantly expanded and updated to include the most recent experimental methodologies and techniques, MS applications in biophysics and structural biology, methods for studying higher order structure and dynamics of proteins, an examination of other biopolymers and synthetic polymers, such as nucleic acids and oligosaccharides, and much more. Featuring high-quality illustrations that illuminate the concepts described in the text, as well as extensive references that enable the reader to pursue further study, Mass Spectrometry in Structural Biology and Biophysics is an indispensable resource for researchers and graduate students working in biophysics, structural biology, protein chemistry, and related fields.

This book is designed to be a central text for young graduate students interested in mass spectrometry as it relates to the study of protein structure and function as well as proteomics. It is a definite must-have work for: - libraries at academic institutions with Master and Graduate programs in biochemistry, molecular biology, structural biology and proteomics - individual laboratories with interests covering these areas - libraries and individual laboratories in the pharmaceutical and biotechnology industries. *Serves as an essential reference to those working in the field *Incorporates the contributions of prominent experts *Features comprehensive coverage and a logical structure

The first systematic summary of biophysical mass spectrometrytechniques Recent advances in mass spectrometry (MS) have pushed the frontiersof analytical chemistry into the biophysical laboratory. As aresult, the biophysical community's acceptance of MS-based methods,used to study protein higher-order structure and dynamics, hasaccelerated the expansion of biophysical MS. Despite this growing trend, until now no single text has presentedthe full array of MS-based experimental techniques and strategiesfor biophysics. Mass Spectrometry in Biophysics expertly closesthis gap in the literature. Covering the theoretical background and technical aspects of eachmethod, this much-needed reference offers an unparalleled overviewof the current state of biophysical MS. Mass Spectrometry inBiophysics begins with a helpful discussion of general biophysicalconcepts and MS-related techniques. Subsequent chaptersaddress: * Modern spectrometric hardware * High-order structure and dynamics as probed by various MS-basedmethods * Techniques used to study structure and behavior of non-nativeprotein states that become populated under denaturingconditions * Kinetic aspects of protein folding and enzyme catalysis * MS-based methods used to extract quantitative information onprotein-ligand interactions * Relation of MS-based techniques to other experimental tools * Biomolecular properties in the gas phase Fully referenced and containing a helpful appendix on the physicsof electrospray mass spectrometry, Mass Spectrometry in Biophysicsalso offers a compelling look at the current challenges facingbiomolecular MS and the potential applications that will likelyshape its future.

Leading practitioners detail revolutionary new spectrometric techniques for the identification and covalent structural characterization of macromolecules, proteins, glycoconjugates, and nucleic acids. Based on the Fourth International Symposium on Mass Spectrometry in the Health and Life Sciences held in San Francisco in 1998, this invaluable book contains tested strategies for solving many significant biomedical research problems. The techniques use mass spectrometry, automated computer processing of spectral information, and gene, protein, and EST databases for genomic and proteomic correlations. Mass Spectrometry in Biology and Medicine offers a unique opportunity to explore and apply these new techniques of mass spectrometry that are revolutionizing the identification and structural characterization of proteins, carbohydrates, and nucleic acids.

Leading practitioners describe in detail advanced methods of mass spectrometry used in structural characterization of biomacromolecules of both natural and recombinant origin. They demonstrate by example how these methodologies can solve a wide array of real-world problems in protein biochemistry, immunology, and glycobiology, as well as for human bacterial pathogens, lipids, and nucleic acids. The book offers a unique opportunity to learn these techniques that are revolutionizing the field. Its authoritative assessment in the context of how to solve important and challenging problems in bioscience and medicine ensures a competitive advantage for today's researchers.

This volume provides an overview of the development and scope of molecular biophysics and in-depth discussions of the major experimental methods that enable biological macromolecules to be studied at atomic resolution. It also reviews the physical chemical concepts that are needed to interpret the experimental results and to understand how the structure, dynamics, and physical properties of biological macromolecules enable them to perform their biological functions. Reviews of research on three disparate biomolecular machines—DNA helicases, ATP synthases, and myosin--illustrate how the combination of theory and experiment leads to new insights and new questions.

The aim of this book is to provide the researcher with important sample preparation strategies in a wide variety of analyte molecules, specimens, methods, and biological applications requiring mass spectrometric analysis as a detection end-point. In this volume we have compiled the contributions from several laboratories which are employing mass spectrometry for biological analysis. With the latest inventions and introduction of highly sophisticated mass spectrometry equipment sample preparation becomes an extremely important bottleneck of biomedical analysis. We have a goal of giving the reader several successful examples of sample preparation, development and optimization, leading to the success in analytical steps and proper conclusions made at the end of the day. This book is structured as a compilation of contributed chapters ranging from protocols to research articles and reviews. The main philosophy of this volume is that sample preparation methods have to be optimized and validated for every project, for every sample type and for every downstream analytical technique.

76 2. Short Oligonucleotide Mass Analysis 76 2. 1. Method Outline 76 2. 2. Design of PCR Primers and Fragments for Analysis 78 2. 3. Typical PCR Reaction Conditions 79 3. Electrospray Ionisation Mass Spectrometry 79 Formation of Ions 3. 1. 79 3. 2. Tandem Mass Spectrometry 79 3. 3. Typical ESI-MS Settings for SOMA 80 4. Purification Procedures 80 4. 1. Phenol/Chloroform Extraction and Ethanol Precipitation 80 4. 2. In-line HPLC Purification 81 5. Genotyping Using SOMA 81 5. 1. APC Genotyping in Human Subjects 81 5. 2. APC Genotyping in Min Mice 85 5. Mutation Detection Using SOMA 86 6. 1. Analysis of p53 Mutations in Liver Cancer Patients 86 6. 1. 1. p53 Mutations in Liver Tumours 87 6. 1. 2. p53 Mutations in Plasma Samples 88 7. Advantages and Disadvantages of SOMA 89 8. Future Perspectives 90 9. Acknowledgements 91 10. References 91 CHAPTER 7 WV. Bienvenut, M. Müller, PM. Palagi, E. Gasteiger, M. Heller, E. Jung, M. Giron, R. Gras, S. Gay, PA. Binz, G J. Hughes, JC. Sanchez, RD. Appel, DF. Hochstrasser Proteomics and Mass Spectrometry: Some Aspects and Recent Developments 1. Introduction to Proteomics 93 2. Protein Biochemical and Chemical Processing Followed by Mass Spectrometric Analysis 94 2. 1. 2-DE Gel Protein Separation 95 Protein Identification Using Peptide Mass Fingerprinting and Robots 96 2. 2. 2. 2. 1. MALDI-MS Analysis 98 2. 2. 2. MS/MS Analysis 102 Improvement of the Identification by Chemical Modification of Peptides 106 2. 2. 3.