New Technologies for Cross-linking Mass Spectrometry

New Technologies for Cross-linking Mass Spectrometry PDF Author: Jared Paul Mohr
Publisher:
ISBN:
Category :
Languages : en
Pages : 0

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Book Description
Cross-linking mass spectrometry is a rapidly evolving technique for obtaining structural information about proteins and protein complexes in their near native state in a high-throughput manner. Cross-linked peptides are challenging to identify due to being low abundance analytes that produce complex fragmentation spectra. While cross-links can provide valuable structural data, these challenges mean that current technologies can sample only a small portion of the protein structures and assemblies that exist in complex systems.In this work I demonstrate three new technologies I developed to enhance cross-linking mass spectrometry experiments. The first section describes the development of the cross-linking search tool, Mango, which enables identification of cross-links in complex samples generated from a variety of cross-linkers. The next section discusses the development of a tetrameric cross-linker and its application in studying the mitochondrial interactome from murine hearts. Higher dimensional cross-linkers move experiments from binary interactions to ternary and quaternary interactions, helping to better characterize interfaces composed of many proteins. The final section describes the development of software to facilitate liquid chromatography coupled to tandem mass spectrometry experiments in a Fourier transform ion cyclotron resonance array mass spectrometer by automating ion selection, ion transfer, and analog signal processing for each cell which allows for parallel acquisition of high-resolution mass spectra. While this hardware has been previously described, modifications to the instrument’s ion handling and analog signal processing enable cross-linking experiments to be carried out with parallel detection and an enhanced duty cycle.

New Technologies for Cross-linking Mass Spectrometry

New Technologies for Cross-linking Mass Spectrometry PDF Author: Jared Paul Mohr
Publisher:
ISBN:
Category :
Languages : en
Pages : 0

Get Book Here

Book Description
Cross-linking mass spectrometry is a rapidly evolving technique for obtaining structural information about proteins and protein complexes in their near native state in a high-throughput manner. Cross-linked peptides are challenging to identify due to being low abundance analytes that produce complex fragmentation spectra. While cross-links can provide valuable structural data, these challenges mean that current technologies can sample only a small portion of the protein structures and assemblies that exist in complex systems.In this work I demonstrate three new technologies I developed to enhance cross-linking mass spectrometry experiments. The first section describes the development of the cross-linking search tool, Mango, which enables identification of cross-links in complex samples generated from a variety of cross-linkers. The next section discusses the development of a tetrameric cross-linker and its application in studying the mitochondrial interactome from murine hearts. Higher dimensional cross-linkers move experiments from binary interactions to ternary and quaternary interactions, helping to better characterize interfaces composed of many proteins. The final section describes the development of software to facilitate liquid chromatography coupled to tandem mass spectrometry experiments in a Fourier transform ion cyclotron resonance array mass spectrometer by automating ion selection, ion transfer, and analog signal processing for each cell which allows for parallel acquisition of high-resolution mass spectra. While this hardware has been previously described, modifications to the instrument’s ion handling and analog signal processing enable cross-linking experiments to be carried out with parallel detection and an enhanced duty cycle.

Proteome Informatics

Proteome Informatics PDF Author: Conrad Bessant
Publisher: Royal Society of Chemistry
ISBN: 1782626735
Category : Science
Languages : en
Pages : 429

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Book Description
The field of proteomics has developed rapidly over the past decade nurturing the need for a detailed introduction to the various informatics topics that underpin the main liquid chromatography tandem mass spectrometry (LC-MS/MS) protocols used for protein identification and quantitation. Proteins are a key component of any biological system, and monitoring proteins using LC-MS/MS proteomics is becoming commonplace in a wide range of biological research areas. However, many researchers treat proteomics software tools as a black box, drawing conclusions from the output of such tools without considering the nuances and limitations of the algorithms on which such software is based. This book seeks to address this situation by bringing together world experts to provide clear explanations of the key algorithms, workflows and analysis frameworks, so that users of proteomics data can be confident that they are using appropriate tools in suitable ways.

Fundamentals of Contemporary Mass Spectrometry

Fundamentals of Contemporary Mass Spectrometry PDF Author: Chhabil Dass
Publisher: John Wiley & Sons
ISBN: 0470118482
Category : Science
Languages : en
Pages : 512

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Book Description
Modern mass spectrometry - the instrumentation and applications in diverse fields Mass spectrometry has played a pivotal role in a variety of scientific disciplines. Today it is an integral part of proteomics and drug discovery process. Fundamentals of Contemporary Mass Spectrometry gives readers a concise and authoritative overview of modern mass spectrometry instrumentation, techniques, and applications, including the latest developments. After an introduction to the history of mass spectrometry and the basic underlying concepts, it covers: Instrumentation, including modes of ionization, condensed phase ionization techniques, mass analysis and ion detection, tandem mass spectrometry, and hyphenated separation techniques Organic and inorganic mass spectrometry Biological mass spectrometry, including the analysis of proteins and peptides, oligosaccharides, lipids, oligonucleotides, and other biological materials Applications to quantitative analysis Based on proven teaching principles, each chapter is complete with a concise overview, highlighted key points, practice exercises, and references to additional resources. Hints and solutions to the exercises are provided in an appendix. To facilitate learning and improve problem-solving skills, several worked-out examples are included. This is a great textbook for graduate students in chemistry, and a robust, practical resource for researchers and scientists, professors, laboratory managers, technicians, and others. It gives scientists in diverse disciplines a practical foundation in modern mass spectrometry.

Functional Proteomics

Functional Proteomics PDF Author: Julie D. Thompson
Publisher: Humana Press
ISBN: 9781588299710
Category : Science
Languages : en
Pages : 0

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Book Description
As the emerging field of proteomics continues to expand at an extremely rapid rate, the relative quantification of proteins, targeted by their function, becomes its greatest challenge. Complex analytical strategies have been designed that allow comparative analysis of large proteomes, as well as in depth detection of the core proteome or the interaction network of a given protein of interest. In Functional Proteomics: Methods and Protocols, expert researchers describe the latest protocols being developed to address the problems encountered in high-throughput proteomics projects, with emphasis on the factors governing the technical choices for given applications. The case studies within the volume focus on the following three crucial aspects of the experimental design: 1) the strategy used for the selection, purification and preparation of the sample to be analyzed by mass spectrometry, 2) the type of mass spectrometer used and the type of data to be obtained from it, and 3) the method used for the interpretation of the mass spectrometry data and the search engine used for the identification of the proteins in the different types of sequence data banks available. As a part of the highly successful Methods in Molecular BiologyTM series, the chapters compile step-by-step, readily reproducible laboratory protocols, lists of the necessary materials and reagents, and tips on troubleshooting and avoiding known pitfalls. Comprehensive and cutting-edge, Functional Proteomics: Methods and Protocols is an ideal resource for all scientists pursuing this developing field and its multitudinous data.

Advancements of Mass Spectrometry in Biomedical Research

Advancements of Mass Spectrometry in Biomedical Research PDF Author: Alisa G. Woods
Publisher: Springer
ISBN: 3319060686
Category : Science
Languages : en
Pages : 601

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Book Description
This volume explores the use of mass spectrometry for biomedical applications. Chapters focus on specific therapeutic areas such as oncology, infectious disease and psychiatry. Additional chapters focus on methodology as well as new technologies and instrumentation. This volume provides readers with a comprehensive and informative manual that will allow them to appreciate mass spectrometry and proteomic research but also to initiate and improve their own work. Thus the book acts as a technical guide but also a conceptual guide to the newest information in this exciting field. Mass spectrometry is the central tool used in proteomic research today and is rapidly becoming indispensable to the biomedical scientist. With the completion of the human genome project and the genomic revolution, the proteomic revolution has followed closely behind. Understanding the human proteome has become critical to basic and clinical biomedical research and holds the promise of providing comprehensive understanding of human physiological processes. In addition, proteomics and mass spectrometry are bringing unprecedented biomarker discovery and are helping to personalize medicine.

Hydrogen Exchange Mass Spectrometry of Proteins

Hydrogen Exchange Mass Spectrometry of Proteins PDF Author: David D. Weis
Publisher: John Wiley & Sons
ISBN: 1118616499
Category : Science
Languages : en
Pages : 422

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Book Description
Hydrogen exchange mass spectrometry is widely recognized for its ability to probe the structure and dynamics of proteins. The application of this technique is becoming widespread due to its versatility for providing structural information about challenging biological macromolecules such as antibodies, flexible proteins and glycoproteins. Although the technique has been around for 25 years, this is the first definitive book devoted entirely to the topic. Hydrogen Exchange Mass Spectrometry of Proteins: Fundamentals, Methods and Applications brings into one comprehensive volume the theory, instrumentation and applications of Hydrogen Exchange Mass Spectrometry (HX-MS) - a technique relevant to bioanalytical chemistry, protein science and pharmaceuticals. The book provides a solid foundation in the basics of the technique and data interpretation to inform readers of current research in the method, and provides illustrative examples of its use in bio- and pharmaceutical chemistry and biophysics In-depth chapters on the fundamental theory of hydrogen exchange, and tutorial chapters on measurement and data analysis provide the essential background for those ready to adopt HX-MS. Expert users may advance their current understanding through chapters on methods including membrane protein analysis, alternative proteases, millisecond hydrogen exchange, top-down mass spectrometry, histidine exchange and method validation. All readers can explore the diversity of HX-MS applications in areas such as ligand binding, membrane proteins, drug discovery, therapeutic protein formulation, biocomparability, and intrinsically disordered proteins.

Mass Spectrometry Analysis for Protein-Protein Interactions and Dynamics

Mass Spectrometry Analysis for Protein-Protein Interactions and Dynamics PDF Author: M. Chance
Publisher: John Wiley & Sons
ISBN: 0470258861
Category : Science
Languages : en
Pages : 325

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Book Description
Presents a wide variety of mass spectrometry methods used to explore structural mechanisms, protein dynamics and interactions between proteins. Preliminary chapters cover mass spectrometry methods for examining proteins and are then followed by chapters devoted to presenting very practical, how-to methods in a detailed way. Includes footprinting and plistex specifically, setting this book apart from the competition.

Quantitative Methods in Proteomics

Quantitative Methods in Proteomics PDF Author: Katrin Marcus
Publisher: Humana Press
ISBN: 9781617798849
Category : Science
Languages : en
Pages : 539

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Book Description
Protein modifications and changes made to them, as well as the quantities of expressed proteins, can define the various functional stages of the cell. Accordingly, perturbations can lead to various diseases and disorders. As a result, it has become paramount to be able to detect and monitor post-translational modifications and to measure the abundance of proteins within the cell with extreme sensitivity. While protein identification is an almost routine requirement nowadays, reliable techniques for quantifying unmodified proteins (including those that escape detection under standard conditions, such as protein isoforms and membrane proteins) is not routine. Quantitative Methods in Proteomics gives a detailed survey of topics and methods on the principles underlying modern protein analysis, from statistical issues when planning proteomics experiments, to gel-based and mass spectrometry-based applications. The quantification of post-translational modifications is also addressed, followed by the “hot” topics of software and data analysis, as well as various overview chapters which provide a comprehensive overview of existing methods in quantitative proteomics. Written in the successful Methods in Molecular BiologyTM series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and easily accessible, Quantitative Methods in Proteomics serves as a comprehensive and competent overview of the important and still growing field of quantitative proteomics.

Ion Mobility-Mass Spectrometry

Ion Mobility-Mass Spectrometry PDF Author: Alison E Ashcroft
Publisher: Royal Society of Chemistry
ISBN: 1839162899
Category : Science
Languages : en
Pages : 621

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Book Description
Over the last decade, the use of ion mobility separation in combination with mass spectrometry analysis has developed significantly. This technique adds a unique extra dimension enabling the in-depth analysis of a wide range of complex samples in the areas of the chemical and biological sciences. Providing a comprehensive guide to the technique, each chapter is written by an internationally recognised expert and with numerous different commercial platforms to choose from, this book will help the end users understand the practicalities of using different instruments for different ion mobility purposes. The first section provides a detailed account of the fundamentals behind the technique and the current range of available instrumentation. The second section focusses on the wide range of applications that have benefitted from ion mobility – mass spectrometry and includes topics taken from current research in the pharmaceutical, metabolomics, glycomics, and structural molecular biology fields. The book is primarily aimed at researchers, appealing to practising chemists and biochemists, as well as those in the pharmaceutical and medical fields.

Integrative Structural Biology with Hybrid Methods

Integrative Structural Biology with Hybrid Methods PDF Author: Haruki Nakamura
Publisher: Springer
ISBN: 9811322007
Category : Science
Languages : en
Pages : 270

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Book Description
This book presents a new emerging concept of "Integrative Structural Biology". It covers current trends of the molecular and cellular structural biology, providing new methods to observe, validate, and keep the structural models of the large cellular machines with recent scientific results. Structures of very large macromolecular machines in cells are being determined by combining observations from complementary experimental methods. Thus, this volume presents the each methods such as X-ray crystallography, NMR spectroscopy, 3DEM, small-angle scattering (SAS), FRET, crosslinking, and enables the readers to understand the hybrid methods. This book discusses how those integrative models should be represented, validated and archived. A unique highlight of this book is discussion of the data validation and archive, which are big problems in this filed along with the progress of this field. The researchers in biology will be interested in this book as a guide book for learning the current structure biology, but also those in structure biology may use this book as a comprehensive reference to cover broad topics.