Download or read book Mass Spectrometry and Hyphenated Techniques in Neuropeptide Research written by Jerzy Silberring and published by John Wiley & Sons. This book was released on 2002-05-02 with total page 576 pages. Available in PDF, EPUB and Kindle. Book excerpt: The first authoritative guide to the application of this vital analytical technique Mass spectrometry is a powerful analytical technique that is used to identify unknown compounds, to quantify known materials, and to elucidate the structural and chemical properties of molecules. In analyzing the effects of experimental drugs on the brain, it is the sole technique for identifying the presence and structure of neuropeptides-substances that indicate the effect of the drug. Mass Spectrometry and Hyphenated Techniques in Neuropeptide Research specifically explains how to apply the technology to this process. Because the book is written by mass spectrometry users, as opposed to mass spectrometrists, the focus remains on practical applications of the technique. The authors demonstrate how mass spectrometry works, how to apply the technique to research, which types of instrumentation should be used for particular requirements, and how to plan experiments. Readers will learn why mass spectrometry provides more outcome features than other techniques in neuropeptide analysis, including simultaneous detection, identification of substances present in mixtures, and sequence information even when the residues are modified, blocked, or unusual. Among the chapters in this comprehensive text are: * Sequencing of Peptides by Nanospray Mass Spectrometry * Laser-Machined Microdevices for Mass Spectrometry * Electron Capture Dissociation of Peptides * Synthesis of Combinatorial Peptide Libraries * Analysis of Tissues That Reflect Nervous System Disease Doctoral students, researchers, and industry professionals in pharmacology, chemistry, biochemistry/biotechnology, and medicine will find Mass Spectrometry and Hyphenated Techniques in Neuropeptide Research to be an indispensable starting point for understanding peptides, their function, and identification.

Download or read book Analysis of Neuropeptides by Liquid Chromatography and Mass Spectrometry written by D.M. Desiderio and published by Elsevier. This book was released on 2000-04-01 with total page 255 pages. Available in PDF, EPUB and Kindle. Book excerpt: Analysis of Neuropeptides by Liquid Chromatography and Mass Spectrometry

Download or read book Mass Spectrometry and Hyphenated Techniques in Neu Ropeptide Research written by Silberring and published by . This book was released on 2002-07-01 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt:

Download or read book Methods in Neurotransmitter and Neuropeptide Research written by S.H. Parvez and published by Elsevier. This book was released on 2013-10-22 with total page 436 pages. Available in PDF, EPUB and Kindle. Book excerpt: This two-part book provides the most advanced biochemical and histochemical analytical methods on neurotransmitters, neuromodulators, and neuropeptides described by international specialists. The biochemical methods include liquid chromatography/electrochemistry, liquid chromatography/fluorescence and chemiluminescence detection, gas chromatography or liquid chromatography with mass spectrometry, and immunoassays. Morphological and physiological methods are also described, including immunohistochemistry, in situ hybridization, in vivo microdialysis, electrophysiological analysis, microwave irradiation, electrophysiological analysis, micro-anatomical dissection, peptide sequence analysis, quantitative autoradiographic analysis, and isolated CNS preparations. The neurotransmitters and neuropeptides include catecholamines, indoleamines, histamine, polyamines, and various neuropeptides, and their related enzymes. The book is the most comprehensive, advanced and versatile volume on the biochemical, morphological and physiological analysis of neurotransmitters and neuropeptides.

Download or read book Analysis of neuropeptides by liquid chromatography and mass spectrometry written by Dominic M. Desiderio and published by . This book was released on 1984 with total page 233 pages. Available in PDF, EPUB and Kindle. Book excerpt:

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.

Download or read book Mass Spectrometry based Neuropeptidomic Study and Functional Discovery of Signaling Peptides written by Yang Liu and published by . This book was released on 2019 with total page 380 pages. Available in PDF, EPUB and Kindle. Book excerpt: Neuropeptides are a group of signaling molecules with great diversity and significant functions in the regulation of physiological processes. The sequence identifications and stress-related studies are highly important in the systematic neuropeptidome characterization of a biological model. Crustaceans are one of the most commonly used animal models in neuropeptide research, providing a simple and well-characterized nervous system. Mass spectrometry (MS) has evolved as a prominent strategy in neuropeptide study, benefiting from its high resolution, high speed and high sensitivity. This dissertation is devoted to the development and application of MS-based techniques in multi-scale characterization of the crustacean neuropeptidome both qualitatively and quantitatively by incorporating various sample collection strategies, sample preparation protocols and MS methods. Collectively, the comprehensive investigation of neuropeptidome documented in this dissertation will lay a solid foundation for future studies of the functional roles of peptide hormones in the crustacean nervous system and the MS-based workflows introduced here will find widespread applications in the studies of signaling molecules utilized by various biological systems.

Download or read book Mass Spectrometry of Peptides written by Dominic M. Desiderio and published by CRC Press. This book was released on 1990-11-14 with total page 438 pages. Available in PDF, EPUB and Kindle. Book excerpt: The purpose of this book is to collect into one volume the research done on the mass spectrometry of peptides. It balances a range of topics including theory, instrumentation, analytical techniques, and biological applications. The scope of the work contains three major sections: ionization methods, instrumental developments, and analysis of peptides. It describes 252Cf plasma desorption and laser-induced multiphoton ionization methodology. This exciting resource covers many new areas, including continuous flow FAB, quantification of human neuropeptides, and peptide mapping. It also discusses Q-FTMS, cross-links, and metal ions.

Download or read book Advancing Mass Spectrometry Methods for Quantification and Characterization of Peptides and Proteins in Complex Biological Samples written by and published by . This book was released on 2015 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: Protein quantification using liquid chromatography mass spectrometry (LC-MS) is a critical component in the discovery and verification of protein candidate biomarkers. Label-free and label-based quantitative methods are both commonly employed, and both have advantages and disadvantages. Label-based methods are capable of analyzing relative protein abundances from multiple samples in one LC-MS experiment, thus enabling high throughput analysis. Unfortunately, the high cost of commercial labels prevents many groups from including them in their proteomics workflows. Our group previously developed a cost-effective isobaric label set, DiLeu, through dimethylation of L-leucine isotopes. The DiLeu reagent performed comparably to commercial options, but multiple improvements and variations to this reagent were needed. For example, the multiplexing capacity of the original DiLeu reagent only permits four quantitative channels, necessitating development of a new label with greater throughput. A portion of this thesis is dedicated to the design, synthesis, and characterization of a new 8-plex DiLeu reagent. This dissertation also recounts the development of isotopic DiLeu (iDiLeu), a 5-plex mass difference variant of DiLeu that allows for researchers to quantify analyte peptide concentrations from a four-point calibration curve in one LC-MS run. One glaring omission in the development of noncommercial isobaric labels is that no protein candidate biomarkers have been discovered by them. Our work rectifies this oversight by identifying several potential biomarkers using custom-synthesized 4-plex DiLeu isobaric tags from the urine of men diagnosed with lower urinary tract symptoms (LUTS). Quantitative strategies presented in this dissertation are complemented by advances in our neuropeptidomics methodologies. In one project, we study the role of protein degradation in crustacean neuropeptidomics and apply a heat stabilization strategy to mitigate the number of protein fragments that interfere with neuropeptide identification. The final project investigates an alternative method to matrix-assisted laser desorption/ionization (MALDI) mass spectrometry imaging (MSI) in crab neuropeptide and lipid imaging. This technique, nanostructure initiator mass spectrometry (NIMS), was unable to ionize neuropeptides in C. borealis brain tissue despite its capability to ionize lipids, showing that sometimes traditional methods like MALDI MSI remain advantageous over more recent but less vetted techniques. The future outlook for our LUTS collaboration and quantitative tag projects is positive because of the new directions still being explored. Overall, the new quantitative techniques developed in this dissertation and the advances in neuropeptide identification will benefit research groups interested in cost-effective label-based quantification methods and neuropeptide characterization in LC-MS and MSI studies.

Download or read book Development of Mass Spectrometric Techniques for the Analysis of Neuropeptides written by Stephanie Shannon Dekeyser and published by . This book was released on 2007 with total page 252 pages. Available in PDF, EPUB and Kindle. Book excerpt:

Download or read book Discovery and Quantification of Crustacean Neuropeptides Using Mass Spectrometry and Informatics Approaches written by Nhu Vu and published by . This book was released on 2022 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: The study of endogenous peptides is important for discovering critical elements involved in many physiological processes and disorders. Neuropeptides are endogenous peptide hormones that regulate these processes by binding to receptors to initiate cellular signaling pathways. Discovery of neuropeptide function is an increasingly popular avenue for disease therapeutics research, resulting in increased need for improved neuropeptide characterization methods. Although mass spectrometry (MS) is well-suited for neuropeptide analysis due to its high sensitivity and capacity for untargeted, global neuropeptide measurements, there are still technical challenges that must be overcome. For example, neuropeptide sequences within a peptide family are highly homologous and therefore must be analyzed in their intact form to obtain high sequence coverage for confident identification and often require high resolution instrumentation. This becomes more challenging as sequence length and post-translational modification complexity increases. In this thesis, I will discuss my efforts in improving MS-based neuropeptidomic workflows, specifically in relation to "top-down" protein and peptide characterization, while highlighting improvements in identification and quantification of peptides through optimized fragmentation and cysteine reactive labeling. I will also discuss my contributions to our group's novel software, HyPep, enabling utility for confident neuropeptide identification. Altogether, these methods have contributed to increased neuropeptide coverage and can be translated to the analysis of other peptide classes.This thesis begins with two reviews of recent developments in MS-based analysis of neuropeptides. Each review outlines examples of how advancements in MS sample preparation, MS acquisition, data analysis, and technology has increased our understanding of neuropeptides, and provides insight on promising directions for further development within each section. My research contributions in Professor Lingjun Li's research group started by applying peptidomics workflows to quantify changes in neuropeptide expression due to hypoxia stress in a crustacean animal model, C. sapidus. Using the same animal model, we also increased the number of detectable neuropeptides using matrix-assisted laser desorption/ionization (MALDI)-MS imaging by incorporating novel tissue wash steps during sample preparation. Over 100 putative peptidomic biomarkers for different levels of hypoxia severity and duration were discovered within multiple tissue types. Four neuropeptide biomarkers were examined for functional activity and two neuropeptides were discovered to increase cardiac output ex vivo, including the first-ever reported function for a crustacean hyperglycemic hormone precursor-related peptide (CPRP). Since CPRP (3.5 kDa) is co-released with crustacean hyperglycemic hormone (CHH) neuropeptide (8.5 kDa), an important neuropeptide that regulates glucose metabolism, we did not detect CHH in these hypoxia studies because its larger size prevented confident detection using typical neuropeptidomic MS workflow and data acquisition methods. Previous members of our research group have developed MS methods to sequence CHH, but limitations of those methods included requiring multiple MS instruments or reliance on targeted MS strategies in which information of potential novel neuropeptides is lost. I built upon those MS acquisition and data analysis methods and was able to sequence novel CHH isoforms and proteoforms, without assistance from genomic or transcriptomic-based sequence prediction, using a Thermo Orbitrap Fusion Lumos instrument. Additionally, I developed novel sample preparation methods to reduce the disulfide bonds and alkylate all six cysteine residues on CHH at high efficiency (83-89% labeling efficiency). While developing methods for CHH are beneficial for analyzing mid-size (8-10 kDa) peptides, I sought to also advance general neuropeptidomics data analysis pipeline. A novel concept for neuropeptide identification software, called HyPep, was created 10 years ago but was not finished and could not be used. To make the software usable, I incorporated features typically used in peptide-spectrum matching (PSM)-based algorithms in proteomics software, such as a false discovery rate. I optimized the software parameters which resulted in HyPep outperforming an analogous commercial software in identifying neuropeptides from four crustacean neuronal tissue types. Collectively, this dissertation is devoted to advancing sample preparation, data acquisition, and data analysis strategies for neuropeptidomics, and these methods are also potentially translatable to analyze other classes of endogenous peptides.

Download or read book Probing Neuropeptidomics by Mass Spectrometry written by and published by . This book was released on 2012 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: In this thesis, an array of biological mass spectrometry (MS) based methods were developed and employed to enable the identification and functional analysis of neuropeptides in the crustacean nervous system. Neuropeptides encompass the largest and most diverse group of signaling molecules in the nervous system. They are necessary for the initiation and regulation of numerous physiological processes such as feeding, reproduction and development. However, the discovery and identification of these important signaling molecules presents unique challenges due to the vast complexity and minute quantities of endogenously expressed peptides. Further functional analysis of neuropeptides needs to be conducted for a comprehensive neuropeptidomic study. Using the decapod crustacean nervous system and associated neuroendocrine organs as experiment model, this dissertation research focuses on the development of an integrated analytical platform for large-scale identification and discovery of neuropeptides, as well as investigation of the functional roles of neuropeptides. The work presented here contributes insight into neuropeptidome of a commercially and scientifically important crustacean species, blue crab /italicCallinectes sapidus/italic and the possible functionality of certain peptides. A multifaceted mass spectrometry approach combining complementary sample preparation techniques, advanced separation strategies, alternative fragmentation methods, mass spectral imaging (MSI) and stable isotopic labeling was used for large-scale identification and to provide comprehensive information on peptide expression level changes under different physiological states. Several new analytical methodologies and tools have been developed to enable enhanced neuropeptidome coverage and improved /italicde novo/italic sequencing. A two dimensional liquid chromatography (2D-LC) method coupling reversed-phase liquid chromatography (RPLC) with nano-LC-ESI-Q-TOF MS/MS was used for neuronal tissue extract analysis and greatly improved the neuropeptidome coverage. A homemade capillary isoelectric focusing (cIEF) system was used for separation of the most acidic neuropeptide family Orcokinins in the sinus gland (SG). Additionally, electron transfer dissociation (ETD) was employed for the identification and sequencing of intact crustacean hyperglycemic hormone precursor related peptides (CPRPs), a group of multiply charged peptides in the SG. More than 150 neuropeptides including over 40 novel ones were identified in blue crab's nervous system. Furthermore, a comparative peptidomic study utilized mass spectral imaging (MSI), stable isotopic labeling and electrophysiological study was developed to provide comprehensive information about peptide expression level changes after food intake. Moreover, a MS-based platform was developed to study the differential degradation of neuropeptides by extracellular peptidases via monitoring degradation rate and degradation products. Collectively, this body of work extends the capability of mass spectrometry as a bioanalytical tool for neuropeptide analysis and advances the basic understanding of neuronal signaling.

Download or read book Monitoring Forest Damage with Mass Spectrometry Based Metabolomics Methods written by Carla Antonio and published by John Wiley & Sons. This book was released on 2024-02-21 with total page 468 pages. Available in PDF, EPUB and Kindle. Book excerpt: Understand forest responses to climate change with this timely introduction Forests are among the most critical parts of our global ecosystem, responsible for the air we breathe, home to most of the earth’s species, and crucial sources of food and raw materials. Forest development is therefore one of the most important areas of ecological study, and damage to forests is potentially existential. Metabolomics, a toolkit which accrues data on interactions between genetic and environmental conditions, promises to advance our understanding of how these vital ecosystems respond to dramatic changes in climate and environment. Monitoring Forest Damage with Mass Spectrometry-Based Metabolomics Methods offers a thorough, accessible discussion of metabolomic techniques and their applications in forest tree research. It promises to enrich the reader’s understanding of how forests are being transformed by globe-spanning changes, and to arm researchers with tools for reacting to these potentially epochal developments. Monitoring Forest Damage with Mass Spectrometry-Based Metabolomics Methods readers will also find: Analysis of specialized secondary metabolites such as phytohormones Detailed discussion of ecologically important tree genera such as Pinus, Populus, Quercus, and many more Supplementary materials related to study design, sample preparation, and instrumental analysis protocols Monitoring Forest Damage with Mass Spectrometry-Based Metabolomics Methods is ideal for researchers in analytical chemistry, mass spectrometry, metabolomics, forest research, the life sciences, and all other related fields.

Download or read book Mass Spectrometry Methods and Applications for Functional Characterization of the Crustacean Neuropeptidome written by Kellen Lynn DeLaney and published by . This book was released on 2020 with total page 524 pages. Available in PDF, EPUB and Kindle. Book excerpt: Neuropeptides are a diverse class of signaling molecules that are responsible for modulating numerous functions within the nervous system. They are challenging to study because, like most signaling molecules, they are present in low abundances in vivo and prone to rapid degradation. In this dissertation, multifaceted mass spectrometry (MS) methods are developed and implemented to characterize neuropeptides within crustacean model organisms. Decapod crustaceans are a useful model for understanding circuit dynamics due to their simplified nervous system, robust central pattern generators, and homology to higher-order organisms, including mammals. This work involves developing methods to improve the coverage of neuropeptides detected by implementing complementary separation and ionization techniques, the identification and reproducibility of neuropeptide analysis with optimized mass spectral acquisition parameters, and the accuracy of sequence information with software specifically designed for neuropeptide analysis. These methods are utilized to profile the neuropeptides present in the cardiac neuromuscular system that may be responsible for modulating heartbeat, as well as feeding-related neuropeptides in tissue and circulating fluid that may be implicated in generating rhythms related to chewing and filtering of food. Overall, this dissertation research develops improved methods for analyzing neuropeptides, demonstrates the utility of these methods for better understanding their functional roles within crustacean model organisms, and provides deeper insight into how these neuropeptides may be involved in modulating key central pattern generators and influencing circuit dynamics.

Download or read book Mass Spectrometry and Stable Isotopes in Nutritional and Pediatric Research written by Henk Schierbeek and published by John Wiley & Sons. This book was released on 2017-01-23 with total page 407 pages. Available in PDF, EPUB and Kindle. Book excerpt: A guide for scientists, pediatricians and students involved in metabolic studies in pediatric research Addresses the availability of modern analytical techniques and how to apply these techniques in metabolic studies Covers the whole range of available mass spectrometric techniques used for metabolic studies including Stable Isotope Methodology Presents the relevance of mass spectrometry and stable isotope methodology in pediatric research covering applications in Nutrition, Obesity, Metabolic Disorders, and Kidney Disorders Focuses on the interactions between nutrients and the endogenous metabolism within the body and how these factors affect the health of a growing infant

Download or read book Mass Spectrometry written by Marek Smoluch and published by John Wiley & Sons. This book was released on 2019-06-17 with total page 361 pages. Available in PDF, EPUB and Kindle. Book excerpt: Provides a comprehensive description of mass spectrometry basics, applications, and perspectives Mass spectrometry is a modern analytical technique, allowing for fast and ultrasensitive detection and identification of chemical species. It can serve for analysis of narcotics, counterfeit medicines, components of explosives, but also in clinical chemistry, forensic research and anti-doping analysis, for identification of clinically relevant molecules as biomarkers of various diseases. This book describes everything readers need to know about mass spectrometry—from the instrumentation to the theory and applications. It looks at all aspects of mass spectrometry, including inorganic, organic, forensic, and biological MS (paying special attention to various methodologies and data interpretation). It also contains a list of key terms for easier and faster understanding of the material by newcomers to the subject and test questions to assist lecturers. Knowing how crucial it is for young researchers to fully understand both the power of mass spectrometry and the importance of other complementary methodologies, Mass Spectrometry: An Applied Approach teaches that it should be used in conjunction with other techniques such as NMR, pharmacological tests, structural identification, molecular biology, in order to reveal the true function(s) of the identified molecule. Provides a description of mass spectrometry basics, applications and perspectives of the technique Oriented to a broad audience with limited or basic knowledge in mass spectrometry instrumentation, theory, and its applications in order to enhance their competence in this field Covers all aspects of mass spectrometry, including inorganic, organic, forensic, and biological MS with special attention to application of various methodologies and data interpretation Includes a list of key terms, and test questions, for easier and faster understanding of the material Mass Spectrometry: An Applied Approach is highly recommended for advanced students, young scientists, and anyone involved in a field that utilizes the technique.

Download or read book Mass Spectrometry for the Analysis of Pesticide Residues and their Metabolites written by Despina Tsipi and published by John Wiley & Sons. This book was released on 2015-05-11 with total page 288 pages. Available in PDF, EPUB and Kindle. Book excerpt: Provides an overview of the use of mass spectrometry (MS) for the analysis of pesticide residues and their metabolites. Presents state of the-art MS techniques for the identification of pesticides and their transformation products in food and environment Covers important advances in MS techniques including MS instrumentation and chromatographic separations (e.g. UPLC, HILIC, comprehensive GCxGC) and applications Illustrates the main sample preparation techniques (SPE, QuEChERS, microextraction) used in combination with MS for the analysis of pesticides Describes various established and new ionization techniques as well as the main MS platforms, software tools and mass spectral libraries