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Microcharacterization of Proteins

Microcharacterization of Proteins
Author: Roland Kellner
Publisher: John Wiley & Sons
Total Pages: 346
Release: 2008-09-26
Genre: Science
ISBN: 3527613978

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Proteomics - the analysis of the whole set of proteins and their functions in a cell - is based on the revolutionary developments which have been achieved in protein analysis during the last years. The number of finished genome projects is growing and in parallel there is a dramatically increasing need to identify the products of revealed genes. Acting on a micro level modern protein chemistry increases our understanding of biological events by elucidating the relevant structure-function relationships. The second edition of the successful title Microcharacterization of Proteins presents a current overview of modern protein analysis: From sample preparation to sequence analysis, mass spectrometry and bioinformatics it informs about the tools needed in protein research. This makes the book indispensable for everyone involved in proteomics!


Methods of Protein Microcharacterization

Methods of Protein Microcharacterization
Author: John E. Shively
Publisher: Springer Science & Business Media
Total Pages: 458
Release: 2008-02-22
Genre: Science
ISBN: 1592594360

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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 Methods in Peptide Mapping for the Characterization of Proteins

New Methods in Peptide Mapping for the Characterization of Proteins
Author: William S. Hancock
Publisher: CRC Press
Total Pages: 280
Release: 1995-10-23
Genre: Medical
ISBN: 9780849378225

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This text is devoted to the characterization of recombinant DNA-derived proteins by peptide mapping. It describes new technological procedures including capillary electrophoresis, analysis of glycopeptides and the use of electrospray and matrix-assisted laser desorption mass spectrometry. The book presents practical procedures for preparing a protein sample, the enzyme digestion, choice of separation method and procedures for the structural analysis of the separated species. Many figures of peptide maps illustrate typical results. Tables of summary information about digestion, separation conditions, and analyses of important protein samples are also presented.


Advanced Technologies for Protein Complex Production and Characterization

Advanced Technologies for Protein Complex Production and Characterization
Author: M. Cristina Vega
Publisher: Springer Nature
Total Pages: 217
Release: 2024
Genre: Biochemistry
ISBN: 3031521935

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Zusammenfassung: This edited volume discusses the identification, discovery, characterization, structure determination and modeling of multicomponent macromolecular complexes, and as such, it fully complements the first volume (ISBN 978-3-319-27214-6), which targeted methods of recombinant production of protein complexes. This book is divided in 8 sections offering a selection of technologies widely used in the characterization of protein and protein-nucleic acid complexes for different purposes and at different scales. From native electrophoresis methods, that are accessible to any reasonably well-equipped laboratory, to the sophisticated setup required for structure determination by cryo-electron microscopy or X-ray crystallography, this book contains a wide variety of clearly explained analytic and preparative approaches, connected with the production techniques developed on the previous volume. The readers will find an integral connection between this book and the first volume, which ensures a comprehensive and updated discussion of the main topics of the discipline. Taken together, these volumes constitute a cohesive and authoritative source of the research on multicomponent macromolecular complexes. In here, we focus on characterization of protein complexes in the broadest sense, which is not typically covered in other sources. Moreover, all chapters are carefully written by world renowned scientists and active researchers, making this volume ideal, not only as a reference source, but also as a companion book for the daily laboratory work. This book is aimed for a wide range of scientists, from science students to experienced researchers, working on protein and protein-nucleic acid complexes, who need a thorough understanding of protein production and complex characterization.


Protein Structure Analysis

Protein Structure Analysis
Author: Roza Maria Kamp
Publisher: Springer Science & Business Media
Total Pages: 340
Release: 1997
Genre: Science
ISBN: 9783540615002

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"Protein Structure Analysis - Preparation and Characterization" is a compilation of practical approaches to the structural analysis of proteins and peptides. Here, about 20 authors describe and comment on techniques for sensitive protein purification and analysis. These methods are used worldwide in biochemical and biotechnical research currently being carried out in pharmaceu tical and biomedical laboratories or protein sequencing facilities. The chapters have been written by scientists with extensive ex perience in these fields, and the practical parts are well documen ted so that the reader should be able to easily reproduce the described techniques. The methods compiled in this book were demonstrated in student courses and in the EMBO Practical Course on "Microsequence Analysis of Proteins" held in Berlin September 10-15, 1995. The topics also derived from a FEBS Workshop, held in Halkidiki, Thessaloniki, Greece, in April, 1995. Most of the authors participated in these courses as lecturers and tutors and made these courses extremely lively and successful. Since polypeptides greatly vary depending on their specific structure and function, strategies for their structural analysis must for the most part be adapted to each individual protein. Therefore, advantages and limitations of the experimen tal approaches are discussed here critically, so that the reader becomes familiar with problems that might be encountered.


Characterization of Proteins

Characterization of Proteins
Author: Felix Franks
Publisher: Springer
Total Pages: 570
Release: 2007-10-03
Genre: Science
ISBN: 1592594379

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Proteins are the servants of life. They occur in all component parts of living organisms and are staggering in their functional var- ty, despite their chemical similarity. Even the simplest single-cell organism contains a thousand different proteins, fulfilling a wide range of life-supporting roles. Their production is controlled by the cell’s genetic machinery, and a malfunction of even one protein in the cell will give rise to pathological symptoms. Additions to the total number of known proteins are constantly being made on an increasing scale through the discovery of mutant strains or their production by genetic manipulation; this latter technology has become known as protein engineering. The in vivo functioning of proteins depends critically on the chemical structure of individual peptide chains, but also on the detailed folding of the chains themselves and on their assembly into larger supramolecular structures. The molecules and their fu- tional assemblies possess a limited in vitro stability. Special methods are required for their intact isolation from the source material and for their analysis, both qualitatively and quantitatively. Proteins are also increasingly used as “industrial components,” e.g., in biosensors and immobilized enzymes, because of their specificity, selectivity, and sensitivity. This requires novel and refined proce- ing methods by which the protein isolate can be converted into a form in which it can be utilized.


Proteome Characterization and Proteomics

Proteome Characterization and Proteomics
Author: Timothy D. Veenstra
Publisher: Academic Press
Total Pages: 445
Release: 2003-10-01
Genre: Science
ISBN: 0080569153

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The content of this volume is designed to reach a wide audience, including those involved with relevant technologies such as electrophoresis and mass spectrometry, to those interested in how proteomics can benefit research. A wide range of techniques are discussed, each specifically designed to address different needs in proteomic analysis. The concluding chapter discusses the important issue related to handling large amounts of data accumulated in proteomic studies. Discusses proteomics in the postgenomic age Includes various strategies for quantitative proteomics Covers the role of MS in structural functional proteomics and proteomics in drug discovery and bioinformatics


Protein Characterization by Capillary Isoelectric Focusing Electrophoresis, Reversed Phase Liquid Chromatography and Mass Spectrometry

Protein Characterization by Capillary Isoelectric Focusing Electrophoresis, Reversed Phase Liquid Chromatography and Mass Spectrometry
Author: Feng Zhou
Publisher: ProQuest
Total Pages:
Release: 2008
Genre: Chromatographic analysis
ISBN: 9780549388937

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With the development of genomics, separation technology and mass spectrometry, proteomics has become an important tool in biological research. Characterization of posttranslational modification (PTM) by mass spectrometry is attracting an increasing attention because of its importance in signal transduction and the difficulty in obtaining this information from genomic methods. Bottom-up methods, which analyze the peptides from enzymatic digestion of proteins, are widely used for proteomic research. However, bottom-up methods may lose some important information when characterizing PTM since not all peptides are detected. In contrast, top-down methods directly characterize intact proteins, which makes it possible to retain PTM information. This dissertation focuses on the characterization of proteins by coupling high throughput methods, capillary isoelectric focusing electrophoresis (CIEF) and electrospray ionization (ESI) mass spectrometry (MS). In this work, a CIEF-RPLC-ESIMS system is designed and constructed for 2-D profiling of protein mixture. The microdialysis membrane based cathodic cell used in this system not only serves as the cathodic cell in CIEF separation, but also transfers the focused proteins separated by CIEF to RPLC. After eliminating the interference of ampholyte, a few fmol of protein can be detected with this system. The detection limit of this system is one order of magnitude lower than 2-D PAGE. By taking the advantage of high resolving power with mass spectrometry and multidimensional separation, the theoretical peak capacity of this system can reach 1x10 6, which is two orders of magnitude higher than theoretical peak capacity of 2-D PAGE. This system is firstly tested with a standard protein mixture. After slight modification, this system is used to profile the intact protein from a yeast enzyme concentrate and a lysate of the green sulfur bacterium Chlorobium tepidum. The pI vs. MW profile obtained from CIEF-RPLC-MS compares favorably with theoretical data derived from C. tepidum genome and experimental data obtained from 2-D-PAGE.


Strategies for Protein Purification and Characterization

Strategies for Protein Purification and Characterization
Author: Daniel R. Marshak
Publisher: CSHL Press
Total Pages: 420
Release: 1996
Genre: Protein engineering
ISBN: 9780879693855

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Cold Spring Harbor Laboratory. Softcover manual of fundamental procedures commonly used in protein biochemistry, for reseachers. Plastic comb spiral binding.