Strategies For Protein And Peptide Characterization And Quantification Using Electron Transfer Dissociation Mass Spectrometry And Intrinsic Fluorescence

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Strategies for Protein and Peptide Characterization and Quantification Using Electron-transfer Dissociation Mass Spectrometry and Intrinsic Fluorescence

Author :
Publisher :
ISBN 13 :
Total Pages : 442 pages
Book Rating : 4.:/5 (811 download)

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Book Synopsis Strategies for Protein and Peptide Characterization and Quantification Using Electron-transfer Dissociation Mass Spectrometry and Intrinsic Fluorescence by :

Download or read book Strategies for Protein and Peptide Characterization and Quantification Using Electron-transfer Dissociation Mass Spectrometry and Intrinsic Fluorescence written by and published by . This book was released on 2012 with total page 442 pages. Available in PDF, EPUB and Kindle. Book excerpt: STRATEGIES FOR PROTEIN AND PEPTIDE CHARACTERIZATION AND QUANTIFICATION USING ELECTRON-TRANSFER DISSOCIATION MASS SPECTROMETRY AND INTRINSIC FLUORESCENCE Jason D. Russell Under the supervision of Associate Professor Joshua J. Coon At the University of Wisconsin-Madison The following chapters detail strategies for peptide and protein sequence analysis featuring electron-transfer dissociation (ETD) tandem mass spectrometry (MS/MS) and quantification using ultraviolet light-induced intrinsic fluorescence (UV-IF). Chapter 1 provides a brief background and overview. Chapter 2 discusses the optimization of the ETD MS/MS duty cycle for large-scale shotgun experiments. In Chapter 3, the first comprehensive analysis of peptide anions using negative ETD (NETD) is detailed. I report in Chapter 4 on the performance of an ion-ion reaction cell for intact protein analysis using large precursor populations for ETD MS/MS. The application of UV-IF for peptide detection and quantification using a custom fluorescence excitation and detection device is discussed in Chapter 5. An analysis of intact proteins from the 26S proteasome of Arabidopsis using top-down mass spectrometry and quantification by UV-IF is described in Chapter 6. In coordination with the Wisconsin Initiative for Science Literacy (WISL), Chapter 7 is a general description of my graduate research intended for non-specialists in an effort to promote science literacy in the broader community.

Protein and Peptide Mass Spectrometry in Drug Discovery

Author : Michael L. Gross
Publisher : John Wiley & Sons
ISBN 13 : 1118116542
Total Pages : 484 pages
Book Rating : 4.1/5 (181 download)

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Book Synopsis Protein and Peptide Mass Spectrometry in Drug Discovery by : Michael L. Gross

Download or read book Protein and Peptide Mass Spectrometry in Drug Discovery written by Michael L. Gross and published by John Wiley & Sons. This book was released on 2011-09-26 with total page 484 pages. Available in PDF, EPUB and Kindle. Book excerpt: The book that highlights mass spectrometry and its application in characterizing proteins and peptides in drug discovery An instrumental analytical method for quantifying the mass and characterization of various samples from small molecules to large proteins, mass spectrometry (MS) has become one of the most widely used techniques for studying proteins and peptides over the last decade. Bringing together the work of experts in academia and industry, Protein and Peptide Mass Spectrometry in Drug Discovery highlights current analytical approaches, industry practices, and modern strategies for the characterization of both peptides and proteins in drug discovery. Illustrating the critical role MS technology plays in characterizing target proteins and protein products, the methods used, ion mobility, and the use of microwave radiation to speed proteolysis, the book also covers important emerging applications for neuroproteomics and antigenic peptides. Placing an emphasis on the pharmaceutical industry, the book stresses practice and applications, presenting real-world examples covering the most recent advances in mass spectrometry, and providing an invaluable resource for pharmaceutical scientists in industry and academia, analytical and bioanalytical chemists, and researchers in protein science and proteomics.

Characterization of Protein Therapeutics using Mass Spectrometry

Author : Guodong Chen
Publisher : Springer Science & Business Media
ISBN 13 : 1441978623
Total Pages : 408 pages
Book Rating : 4.4/5 (419 download)

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Book Synopsis Characterization of Protein Therapeutics using Mass Spectrometry by : Guodong Chen

Download or read book Characterization of Protein Therapeutics using Mass Spectrometry written by Guodong Chen and published by Springer Science & Business Media. This book was released on 2014-07-08 with total page 408 pages. Available in PDF, EPUB and Kindle. Book excerpt: This book highlights current approaches and future trends in the use of mass spectrometry to characterize protein therapies. As one of the most frequently utilized analytical techniques in pharmaceutical research and development, mass spectrometry has been widely used in the characterization of protein therapeutics due to its analytical sensitivity, selectivity, and specificity. This book begins with an overview of mass spectrometry techniques as related to the analysis of protein therapeutics, structural identification strategies, quantitative approaches, followed by studies involving characterization of process related protein drug impurities/degradants, metabolites, higher order structures of protein therapeutics. Both general practitioners in pharmaceutical research and specialists in analytical sciences will benefit from this book that details step-by-step approaches and new strategies to solve challenging problems related to protein therapeutics research and development.

Protein Analysis using Mass Spectrometry

Author : Mike S. Lee
Publisher : John Wiley & Sons
ISBN 13 : 1119359368
Total Pages : 282 pages
Book Rating : 4.1/5 (193 download)

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Book Synopsis Protein Analysis using Mass Spectrometry by : Mike S. Lee

Download or read book Protein Analysis using Mass Spectrometry written by Mike S. Lee and published by John Wiley & Sons. This book was released on 2017-05-26 with total page 282 pages. Available in PDF, EPUB and Kindle. Book excerpt: Presents Practical Applications of Mass Spectrometry for Protein Analysis and Covers Their Impact on Accelerating Drug Discovery and Development Covers both qualitative and quantitative aspects of Mass Spectrometry protein analysis in drug discovery Principles, Instrumentation, Technologies topics include MS of peptides, proteins, and ADCs , instrumentation in protein analysis, nanospray technology in MS protein analysis, and automation in MS protein analysis Details emerging areas from drug monitoring to patient care such as Identification and validation of biomarkers for cancer, targeted MS approaches for biomarker validation, biomarker discovery, and regulatory perspectives Brings together the most current advances in the mass spectrometry technology and related method in protein analysis

Mass Spectrometry of Proteins and Peptides

Author : Mary S. Lipton
Publisher : Humana Press
ISBN 13 : 9781627037969
Total Pages : 0 pages
Book Rating : 4.0/5 (379 download)

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Book Synopsis Mass Spectrometry of Proteins and Peptides by : Mary S. Lipton

Download or read book Mass Spectrometry of Proteins and Peptides written by Mary S. Lipton and published by Humana Press. This book was released on 2014-11-27 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: When the last edition of this book was published in 2000, the field of proteomics was in its infancy. At that time, multidimensional liquid chromatographic separations were being introduced as an alternative to traditional gel-based techniques for separating complex protein and peptide mixtures prior to mass spectrometric detection. Today, this approach – referred to as shotgun proteomics – is considered routine for lar- scale global analyses of protein mixtures. Now in its adolescence, proteomics is fundamentally transforming biological and medical research. Much of this transformation can be attributed to technological advancements, particularly in mass spectrometry. Much wider accessibility of hi- resolution and mass measurement accuracy instrumentation in recent years has ini- ated a new revolution in the field by providing more reliable data and shifting the focus from cataloging proteins to precisely quantifying changes in protein abundance over time and in response to stimuli. Advanced mass spectrometers and novel ion d- sociation schemes such as electron transfer/capture dissociation make it possible to venture boldly into the maze of protein posttranslational modifications, which are an integral component of understanding functional proteomics in the spatial and t- poral domains. Another area that has benefited from these advancements is top-down proteomics, an emerging method essential for characterizing various protein variants that has potentially high impact in biomedical research.

Advancing Electron Transfer Dissociation Technologies for Characterization of Proteomes and Post-translational Modifications

Author : Nicholas M. Riley
Publisher :
ISBN 13 :
Total Pages : 0 pages
Book Rating : 4.:/5 (112 download)

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Book Synopsis Advancing Electron Transfer Dissociation Technologies for Characterization of Proteomes and Post-translational Modifications by : Nicholas M. Riley

Download or read book Advancing Electron Transfer Dissociation Technologies for Characterization of Proteomes and Post-translational Modifications written by Nicholas M. Riley and published by . This book was released on 2018 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: This dissertation presents research focusing on the development of new instrumentation and methodology to leverage ion-ion reactions for proteomic analyses. Electron transfer dissociation (ETD) technologies have proven a valuable alternative to collision-based fragmentation methods for sequencing peptides and proteins to advance global proteome characterization. Chapter 1 outlines the core concepts central to mass spectrometry (MS)-based proteomics, in addition to the basic principles of ETD and various strategies to improve its efficacy - including the technology that is the focus of this work, i.e., activated ion ETD (AI-ETD). Chapter 2 describes the first application of AI-ETD to intact proteins, which are more chemically complex and, thus, more difficult to sequence, than their peptide counterparts. Chapter 3 discusses a new strategy to improve signalto- noise in ETD spectra, which is especially beneficial for intact protein analysis and which has been incorporated into the newest generation of commercially available quadrupole-Orbitrap-linear ion trap hybrid MS systems. AI-ETD capabilities were also recently implemented on this stateof- the-art MS system (Chapter 4), and the ability to perform AI-ETD on this instrument enables comprehensive sequence coverage of moderately-sized intact proteins (Chapter 5), significantly improves proteoform characterization in large-scale analyses of complex mixtures of intact proteins (Chapter 6), and also enhances characterization of larger intact proteins (Chapter 7). Furthermore, AI-ETD improves characterization of post-translational modifications. Chapter 8 demonstrates the utility of AI-ETD for phosphosite localization in phosphopeptides and intact phosphoproteins, and Chapter 9 presents the largest glycoproteomic study to date by using AI-ETD to interrogate intact N-glycopeptides. Beyond positive-mode analyses of peptide and protein cations, ion-ion reactions also bring unique benefits to negative-mode analyses of precursor anions, where collision-based dissociation fails to consistently produce sequence-informative fragments. Chapter 10 describes implementation of negative ETD (NETD) and activated ion NETD (AI-NETD) and their application to whole-proteome sequencing in the negative mode, and Chapter 11 presents a modified search algorithm to improve interpretation of large-scale NETD and AI-NETD data. Conclusions and future directions of these projects are discussed in Chapter 12.

Analysis of Protein Post-Translational Modifications by Mass Spectrometry

Author : John R. Griffiths
Publisher : John Wiley & Sons
ISBN 13 : 1119045851
Total Pages : 414 pages
Book Rating : 4.1/5 (19 download)

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Book Synopsis Analysis of Protein Post-Translational Modifications by Mass Spectrometry by : John R. Griffiths

Download or read book Analysis of Protein Post-Translational Modifications by Mass Spectrometry written by John R. Griffiths and published by John Wiley & Sons. This book was released on 2016-11-07 with total page 414 pages. Available in PDF, EPUB and Kindle. Book excerpt: Covers all major modifications, including phosphorylation, glycosylation, acetylation, ubiquitination, sulfonation and and glycation Discussion of the chemistry behind each modification, along with key methods and references Contributions from some of the leading researchers in the field A valuable reference source for all laboratories undertaking proteomics, mass spectrometry and post-translational modification research

New Methods in Peptide Mapping for the Characterization of Proteins

Author : William S. Hancock
Publisher : CRC Press
ISBN 13 : 9780849378225
Total Pages : 280 pages
Book Rating : 4.3/5 (782 download)

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Book Synopsis New Methods in Peptide Mapping for the Characterization of Proteins by : William S. Hancock

Download or read book New Methods in Peptide Mapping for the Characterization of Proteins written by William S. Hancock and published by CRC Press. This book was released on 1995-10-23 with total page 280 pages. Available in PDF, EPUB and Kindle. Book excerpt: 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.

Investigation of Energy Transfer, Quantification, and Localization of Peptides and Proteins by Fluorescence Spectroscopy and Mass Spectrometry

Download Investigation of Energy Transfer, Quantification, and Localization of Peptides and Proteins by Fluorescence Spectroscopy and Mass Spectrometry PDF Online Free

Author : Suraj Saraswat
Publisher :
ISBN 13 :
Total Pages : 138 pages
Book Rating : 4.:/5 (812 download)

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Book Synopsis Investigation of Energy Transfer, Quantification, and Localization of Peptides and Proteins by Fluorescence Spectroscopy and Mass Spectrometry by : Suraj Saraswat

Download or read book Investigation of Energy Transfer, Quantification, and Localization of Peptides and Proteins by Fluorescence Spectroscopy and Mass Spectrometry written by Suraj Saraswat and published by . This book was released on 2012 with total page 138 pages. Available in PDF, EPUB and Kindle. Book excerpt: Peptides and proteins are integral biomolecules found in all organisms, and are responsible for movement, molecular transport, catalysis, and energy transfer. This dissertation is aimed on the analysis of energy transfer involving fluorescent proteins and development of methodologies for quantitative and imaging analyses of proteins. Such studies are important because energy transfer plays a significant role in numerous chemical, physical, and biological processes, while quantity and location of peptides and proteins are often closely associated with their functions. In the initial study, energy transfer from fluorescent phycobiliproteins to noble metal nanoparticles was analyzed. Solutions of highly fluorescent phycobiliprotein B-phycoerythrin (B-PE) were mixed with colloidal Au and Ag nanoparticles and were characterized by steady-state and time-resolved fluorescence spectroscopy to determine the magnitude and mechanism of the energy transfer. It was found that the protein fluorescence was quenched after the addition of metal nanoparticles. Electron microscopy and absorption spectroscopy confirmed that B-PE was adsorbed onto the nanoparticles, creating a favorable geometry for quenching. Time-resolved fluorescence spectroscopy showed that B-PE fluorescence lifetimes decreased from 2.2 ns to 0.5 and 0.6 ns upon adsorption onto Au and Ag nanoparticles, respectively, corresponding to energy transfer efficiencies of>70%. Our results, which include lifetimes, efficiencies, and energy transfer distances, show that energy was transferred via the surface energy transfer (SET) mechanism, rather than Forster resonance energy transfer (FRET). The results also imply that efficient energy transfer between proteins and metal nanoparticles may be possible regardless of whether or not resonance conditions between emission spectra of the proteins and absorption spectra of nanoparticles are achieved. The second project dealt with quantitative analysis of proteins by a combination of fluorescence spectroscopy and electrospray ionization (ESI) mass spectrometry (MS). While MS is a very sensitive and versatile detection technique, quantification of compounds by MS is challenging due to relatively low reproducibility of ionization. We investigated the native fluorescence of phenylalanine, tryptophan, and tyrosine as a tool that can improve quantification of peptides and proteins by LC-ESI-MS. Natively fluorescent amino acids as well as peptides and proteins containing them were successfully separated by HPLC, and quantified with a spectrofluorimetric detector and a mass spectrometer. Two detectors connected in series enabled sequential measurements of fluorescence intensities and ion signals as well as structural characterization of separated polypeptides. Fluorescence detector provided better linearity and repeatability of quantification than mass spectrometer, and similar sensitivity for most of biomolecules analyzed. The fluorescence signal was linear over 3-4 orders of magnitude with limit of detection in the low picomole or high femtomole range. Hence, spectrofluorimetric detection of native fluorescence can be used as a reliable method that can facilitate quantification of peptides and proteins by LC-ESI-MS. In addition to analyzing native fluorescence of peptides and proteins, the effect of fluorescence dye labeling on ESI efficiency and charging of polypeptides was studied. Peptides and proteins were labeled with fluorescein isothiocyanate (FITC) and separated from the dye by size exclusion chromatography and HPLC. The ESI-MS and LC-ESI-MS analyses of mixtures of labeled and unlabeled polypeptides confirmed that labeling was successful, leading to slight increase in the charging of the most abundant ions in the mass spectra. Further experiments will confirm how derivatization of the peptides and proteins by FITC affects their ionization efficiencies. In the last project, distribution and localization of biomolecules within the mouse tissue sections was studied by matrix-assisted laser desorption/ionization (MALDI)-imaging mass spectrometry (IMS). Peptides and proteins were imaged in brain and spinal cord tissue sections originating from non-transgenic mice and transgenic mice containing the gene for amyotrophic lateral sclerosis (ALS). Tissue sections from apparently healthy and diseased (ALS) mice were coated with sinapinic acid (SA) or a-cyano-4-hydroxycinnamic acid (CHCA) matrixes and imaged using a MALDI-MS instrument. These imaging experiments enabled determination of distribution and localization of peptides and proteins in the spinal cord and brain tissue sections.

Mass Spectrometry-based Strategies for Protein Biophysics

Author : Richard Yu-Cheng Huang
Publisher :
ISBN 13 :
Total Pages : 548 pages
Book Rating : 4.:/5 (812 download)

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Book Synopsis Mass Spectrometry-based Strategies for Protein Biophysics by : Richard Yu-Cheng Huang

Download or read book Mass Spectrometry-based Strategies for Protein Biophysics written by Richard Yu-Cheng Huang and published by . This book was released on 2012 with total page 548 pages. Available in PDF, EPUB and Kindle. Book excerpt: Two important biophysical characteristics of proteins, their interaction with ligands and their post-translational modifications, modulate various biological processes including signal transduction, chemical synthesis, and cell function. Protein-ligand interactions include the interactions with protein, peptide, DNA, and metal. Characterization of the physical properties of these interactions (binding interfaces, binding affinities, and the protein conformational changes due to the binding) is essential in understanding the mechanism of related diseases and, more importantly, in future drug design. Mass spectrometry, with its own revolution and improvement, becomes a powerful tool in protein and peptide analysis. In this thesis, we applied two mass spectrometry-based strategies, proteomics and protein footprinting, to characterize these biophysical properties of three disease-related proteins, connexin 43 (Cx43), troponin, and apolipoprotein E (ApoE), and Fenna-Matthews-Olson protein (FMO), which is the key factor in energy transfer of the photosynthetic system of green sulfur bacteria. By the combination of standard proteomics workflow and two fragmentation methods, collision-induced dissociation (CID) and electron transfer dissociation (ETD), we successfully identified 15 serine residues, including one novel site, in the Cx43-CT that are phosphorylated by CaMKII, the activity of which may be important in regulating Cx43 in normal and diseased hearts. We further utilized hydrogen/deuterium exchange (H/DX), one mass spectrometry-based protein footprinting strategy, to examine the binding affinities of troponin C (TnC), a cardiac disease-related protein, with its four metal binding ligands (Ca2+), and their binding order. We then expanded this approach to elucidate the dynamics of TnC within the complex and its interactions with other subunits (TnT and TnI) at peptide-level resolution. This same approach was also applied to two protein-ligand complexes: (1) the interaction of FMO and its binding partner, CsmA baseplate protein, in which the orientation of FMO between chlorosome and membrane can be confirmed, and (2) the interaction of ApoE and Abeta 40, which are both key factors in Alzheimer's disease. Moreover, we improved the spatial resolution of H/DX to residue-level by conducting ETD fragmentation in the study of ApoE oligomerization. Our results reveal, for the first time, the amino acid residues involved in its self-oligomerization. These six applications of mass spectrometry-based approaches show their potential in the characterization of different protein biophysical properties. The investigation of a more complex protein system can then be pursued.

The Development and Implementation of Mass Spectrometry Methods for the Characterization of Proteins and Metabolites

Author : Matthew Rush
Publisher :
ISBN 13 :
Total Pages : 0 pages
Book Rating : 4.:/5 (11 download)

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Book Synopsis The Development and Implementation of Mass Spectrometry Methods for the Characterization of Proteins and Metabolites by : Matthew Rush

Download or read book The Development and Implementation of Mass Spectrometry Methods for the Characterization of Proteins and Metabolites written by Matthew Rush and published by . This book was released on 2018 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: The work described in this dissertation involves the development and application of mass spectrometry methods for the analysis of peptides, proteins, and metabolites. Chapter 1 gives a brief overview of the history and fundamentals of bioanalytical mass spectrometry, the utility of electron transfer dissociation (ETD) for the analysis of peptides and proteins, and the use of high resolution gas-chromatography mass spectrometry (GC-MS) to characterize volatile small molecule metabolites from complex mixtures. Chapter 2 explores the role of the reagent cation in negative electron transfer dissociation (NETD) for the analysis of peptide anions. Then Chapter 3 demonstrates the use of activated-ion NETD (AI-NETD) to analyze the negative-mode proteome of yeast. This work proved to generate far greater depth of coverage than previous negative-mode experiments, owing largely to the introduction of IR radiation concurrent to the NETD reaction, which proved to drastically improve peptide identification. For this study, a high pH chromatography method was also developed to greatly improve the ionization of peptide anions using negative electrospray ionization. Chapter 4 characterizes activated-ion ETD (AI-ETD) as a fragmentation method to interrogate proteins with intact disulfide bonds. This method greatly improved the sequence coverage and sequence ion generation compared to other commonly used fragmentation techniques for a set standard proteins. Chapter 5 outlines the creation of a high resolution metabolite mass spectral library using a Q Exactive GC mass spectrometer. This library is then employed to identify metabolites from yeast and human cell cultures, significantly improving the identification confidence over commercially available spectral libraries. Lastly, Chapter 6 describes the multi-omic analysis of yeast strains with a single gene deletion, aiming to correlate genes of known function with genes unknown function.

Enhanced Protein Characterization Through Selective Derivatization and Electrospray Ionization Tandem Mass Spectrometry

Author : Lisa Anne Vasicek
Publisher :
ISBN 13 :
Total Pages : 338 pages
Book Rating : 4.:/5 (759 download)

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Book Synopsis Enhanced Protein Characterization Through Selective Derivatization and Electrospray Ionization Tandem Mass Spectrometry by : Lisa Anne Vasicek

Download or read book Enhanced Protein Characterization Through Selective Derivatization and Electrospray Ionization Tandem Mass Spectrometry written by Lisa Anne Vasicek and published by . This book was released on 2011 with total page 338 pages. Available in PDF, EPUB and Kindle. Book excerpt: There continue to be great strides in the field of proteomics but as samples become more complex, the ability to increase sequence coverage and confidence in the identification becomes more important. Several methods of derivatization have been developed that can be used in combination with tandem mass spectrometry to identify and characterize proteins. Three types of activation, including infrared multiphoton dissociation, ultraviolet photodissociation, and electron transfer dissociation, are enhanced in this dissertation and compared to the conventional method of collisional induced dissociation (CID) to demonstrate the improved characterization of proteins. A free amine reactive phosphate group was synthesized and used to modify the N-terminus of digested peptides. This phosphate group absorbs at the IR wavelength of 10.6 [mu]m as well as the Vacuum-ultraviolet (VUV) due to an aromatic group allowing modified peptides to be dissociated by infrared multi-photon dissociation (IRMPD) or ultraviolet photodissociation (UVPD) whereas peptides without this chromophore are less responsive to IR or UV irradiation. The PD spectra for these modified peptides yield simplified MS/MS spectra due to the neutralization of all N-terminal product ions from the incorporation the negatively charged phosphate moiety. This is especially advantageous for UVPD due to the great number of product ions produced due to the higher energy deposition of the UV photons. The MS/MS spectra also produce higher sequence coverage in comparison to CID of the modified or unmodified peptides due to more informative fragmentation pathways generated upon PD from secondary dissociation and an increased ion trapping mass range. IRMPD is also implemented for the first time on an orbitrap mass spectrometer to achieve high resolution analysis of IR chromophore-derivatized samples as well as top-down analysis of unmodified proteins. High resolution/high mass accuracy analysis is extremely beneficial for characterization of complex samples due to the likelihood of false positives at lower resolutions/accuracies. For electron transfer dissociation, precursor ions in higher charge states undergo more exothermic electron transfer and thus minimize non-dissociative charge reduction. In this dissertation, cysteine side chains are alkylated with a fixed charge to deliberately increase the charge states of peptides and improve electron transfer dissociation. ETD can also be used to study protein structure by derivatizing the intact structure with a hydrazone reagent. A hydrazone bond will be preferentially cleaved during ETD facilitating the recognition of any modified residues through a distinguishing ETD fragmentation spectrum.

Protein Structure Analysis

Author : Roza Maria Kamp
Publisher : Springer Science & Business Media
ISBN 13 : 9783540615002
Total Pages : 340 pages
Book Rating : 4.6/5 (15 download)

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Book Synopsis Protein Structure Analysis by : Roza Maria Kamp

Download or read book Protein Structure Analysis written by Roza Maria Kamp and published by Springer Science & Business Media. This book was released on 1997 with total page 340 pages. Available in PDF, EPUB and Kindle. Book excerpt: "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.

Mass Spectrometry of Proteins and Peptides

Author : Mary S. Lipton
Publisher : Humana Press
ISBN 13 : 9781934115480
Total Pages : 470 pages
Book Rating : 4.1/5 (154 download)

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Book Synopsis Mass Spectrometry of Proteins and Peptides by : Mary S. Lipton

Download or read book Mass Spectrometry of Proteins and Peptides written by Mary S. Lipton and published by Humana Press. This book was released on 2008-12-18 with total page 470 pages. Available in PDF, EPUB and Kindle. Book excerpt: When the last edition of this book was published in 2000, the field of proteomics was in its infancy. At that time, multidimensional liquid chromatographic separations were being introduced as an alternative to traditional gel-based techniques for separating complex protein and peptide mixtures prior to mass spectrometric detection. Today, this approach – referred to as shotgun proteomics – is considered routine for lar- scale global analyses of protein mixtures. Now in its adolescence, proteomics is fundamentally transforming biological and medical research. Much of this transformation can be attributed to technological advancements, particularly in mass spectrometry. Much wider accessibility of hi- resolution and mass measurement accuracy instrumentation in recent years has ini- ated a new revolution in the field by providing more reliable data and shifting the focus from cataloging proteins to precisely quantifying changes in protein abundance over time and in response to stimuli. Advanced mass spectrometers and novel ion d- sociation schemes such as electron transfer/capture dissociation make it possible to venture boldly into the maze of protein posttranslational modifications, which are an integral component of understanding functional proteomics in the spatial and t- poral domains. Another area that has benefited from these advancements is top-down proteomics, an emerging method essential for characterizing various protein variants that has potentially high impact in biomedical research.

Instrumental Analysis of Intrinsically Disordered Proteins

Author : Vladimir Uversky
Publisher : John Wiley & Sons
ISBN 13 : 0470602600
Total Pages : 792 pages
Book Rating : 4.4/5 (76 download)

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Book Synopsis Instrumental Analysis of Intrinsically Disordered Proteins by : Vladimir Uversky

Download or read book Instrumental Analysis of Intrinsically Disordered Proteins written by Vladimir Uversky and published by John Wiley & Sons. This book was released on 2011-01-31 with total page 792 pages. Available in PDF, EPUB and Kindle. Book excerpt: Instrumental techniques for analyzing intrinsically disordered proteins The recently recognized phenomenon of protein intrinsic disorder is gaining significant interest among researchers, especially as the number of proteins and protein domains that have been shown to be intrinsically disordered rapidly grows. The first reference to tackle this little-documented area, Instrumental Analysis of Intrinsically Disordered Proteins: Assessing Structure and Conformation provides researchers with a much-needed, comprehensive summary of recent achievements in the methods for structural characterization of intrinsically disordered proteins (IDPs). Chapters discuss: Assessment of IDPs in the living cell Spectroscopic techniques for the analysis of IDPs, including NMR and EPR spectroscopies, FTIR, circular dichroism, fluorescence spectroscopy, vibrational methods, and single-molecule analysis Single-molecule techniques applied to the study of IDPs Assessment of IDP size and shape Tools for the analysis of IDP conformational stability Mass spectrometry Approaches for expression and purification of IDPs With contributions from an international selection of leading researchers, Instrumental Analysis of Intrinsically Disordered Proteins: Assessing Structure and Conformation fills an important need in a rapidly growing field. It is required reading for biochemists, biophysicists, molecular biologists, geneticists, cell biologists, physiologists, and specialists in drug design and development, proteomics, and molecular medicine with an interest in proteins and peptides.

Techniques in Protein Chemistry

Author : John W. Crabb
Publisher : Elsevier
ISBN 13 : 1483217590
Total Pages : 591 pages
Book Rating : 4.4/5 (832 download)

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Book Synopsis Techniques in Protein Chemistry by : John W. Crabb

Download or read book Techniques in Protein Chemistry written by John W. Crabb and published by Elsevier. This book was released on 2014-06-28 with total page 591 pages. Available in PDF, EPUB and Kindle. Book excerpt: Techniques in Protein Chemistry V highlights current methods in peptide and protein mass spectrometry, sequence and amino acid analysis, fragmentations, separations, protein folding and modeling, peptide and protein NMR, and peptide synthesis. This volume emerged from the manuscripts presented at the Seventh Symposium of the Protein Society, held in San Diego on July 24-28, 1993. This volume is organized into eight parts encompassing 61 chapters. The first part surveys the peptide and protein characterization, detection, and analysis by mass spectrometry. The subsequent parts describe the structural characterization and analysis of posttranslational processing events, as well as the characterization of protein and amino acid sequences using several analytical techniques. Other parts explore other analytical methods for peptide and protein separations; some aspects involved in protein design and functional domain analysis; and the evaluation of protein conformation, folding, and modeling. The last parts contain research papers on NMR analysis of peptide and protein solution structures. These parts also look into topics related to peptide synthesis and peptide libraries. This book is intended primarily for protein and analytical chemists.

Improvements to the Identification and Quantification of Peptides and Proteins

Author : Christopher Michael Rose
Publisher :
ISBN 13 :
Total Pages : 294 pages
Book Rating : 4.:/5 (962 download)

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Book Synopsis Improvements to the Identification and Quantification of Peptides and Proteins by : Christopher Michael Rose

Download or read book Improvements to the Identification and Quantification of Peptides and Proteins written by Christopher Michael Rose and published by . This book was released on 2014 with total page 294 pages. Available in PDF, EPUB and Kindle. Book excerpt: The following chapters contain a plethora of proteomic techniques aimed at the improvement of methods for the identification and quantification of peptide and protein species. The genesis of mass spectrometry for the analysis of biological molecules and the advances that have resulted in the large-scale identification and quantification of proteins are detailed in Chapter 1. Chapter 2 discusses the application of quantitative proteomic techniques for the large-scale analysis of a biological system, Medicago truncatula. The third chapter discusses the implementation of an online algorithm to identify peptides in real time (InSeq), enabling alterations to the instrument method that increase the quantitative accuracy of measurements or aid in post translational modification localization. Chapters 4 to 6 outline advances to the implementation of electron transfer dissociation aimed at increasing the ability to identify protein (Chapter 4) or peptide (Chapters 5 and 6) species. The remainder of this thesis details the application of neutron encoded (NeuCode) mass labels to peptide identification without tandem MS (Chapter 7), top-down quantification of intact proteoforms (Chapter 8), labeling of mammals that enables multiplexed quantitative analysis of mammalian tissue after only 10 days of labeling time (Chapter 9), and targeted quantification of more than 1,000 peptides (Chapter 10). Conclusions and future directions relating to the content of this thesis are discussed in Chapter 11.