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 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 0 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 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 Exploring Neuropeptides in the Brain Using Capillary Liquid Chromatography Matrix Assisted Laser Desorption Ionization Mass Spectrometry written by Hui Wei and published by . This book was released on 2006 with total page 182 pages. Available in PDF, EPUB and Kindle. Book excerpt: An endogenous neuropeptide, orphanin FQ (N/OFQ, was characterized by combining the behavioral studies and quantification methods utilizing stable isotope incorporated standard in MALDI-TOF-MS. N/OFQ was found to produce antinociceptive effects in monkeys without pruritus.

Download or read book Peptidomics written by Mikhail Soloviev and published by John Wiley & Sons. This book was released on 2007-12-21 with total page 432 pages. Available in PDF, EPUB and Kindle. Book excerpt: The definitive guide to peptidomics- a hands-on lab reference The first truly comprehensive book about peptidomics for protein and peptide analysis, this reference provides a detailed description of the hows and whys of peptidomics and how the techniques have evolved. With chapters contributed by leading experts, it covers naturally occurring peptides, peptidomics methods and new developments, and the peptidomics approach to biomarker discovery. Explaining both the principles and the applications, Peptidomics: Methods and Applications: * Features examples of applications in diverse fields, including pharmaceutical science, toxicity biomarkers, and neuroscience * Details the successful peptidomic analyses of biological material ranging from plants to mammals * Describes a cross section of analytical techniques, including traditional methodologies, emerging trends, and new techniques for high throughput approaches An enlightening reference for experienced professionals, this book is sufficiently detailed to serve as a step-by-step guide for beginning researchers and an excellent resource for students taking biotechnology and proteomics courses. It is an invaluable reference for protein chemists and biochemists, professionals and researchers in drug and biopharmaceutical development, analytical and bioanalytical chemists, toxicologists, and others.

Download or read book Probing Neuropeptidomics by Mass Spectrometry written by and published by . This book was released on 2012 with total page 548 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 Liquid Chromatography mass Spectrometry based Neuropeptide Characterizations Across Neuronal Models written by and published by . This book was released on 2012 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt:

Download or read book Liquid Chromatography written by K.E. Bosse and published by Elsevier Inc. Chapters. This book was released on 2013-01-08 with total page 86 pages. Available in PDF, EPUB and Kindle. Book excerpt: To achieve quantitative measurements of neurotransmitters, researchers employ analytical methods based on balancing the amenability and limitations of a technique for a specific application. In particular, in-vivo microdialysis sampling has proven indispensable toward improving our understanding of the biological roles mediated by diverse neuroactive compounds. This chapter discusses the development and application of liquid-chromatography methods for analysis of a broad range of neurotransmitters and their metabolites from tissue and dialysate samples in both experimental and clinical settings. Liquid chromatography–based methods are capable of resolving neurotransmitters from complex biological matrices and offer flexibility in coupling to various detection techniques, such as mass spectrometry, electrochemical, and fluorescence. Improved column technologies and separation conditions offer increased efficiency for chromatographic resolution, decreased separation time, improved temporal resolution, and enhanced multianalyte capability. Together, microdialysis sampling and liquid chromatography–based methods continue to be principle techniques widely employed to examine neurotransmitter function in the central nervous system.

Download or read book Advances in Capillary Liquid Chromatography written by Holly Merideth Shackman and published by . This book was released on 2005 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt:

Download or read book Neuropeptidomics MS base Peptide Identification and Quantitation Study written by Ping Yin and published by . This book was released on 2011 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt: Neuropeptidomics refers to a global characterization approach for the investigation of neuropeptides, often under specific physiological conditions. Neuropeptides comprise a complex set of cell-to-cell signaling molecules that are important in regulatory functions and behavior in the nervous system. Neuropeptide production typically involves endoproteolytic prohormone cleavage, exoproteolytic cleavage, and post-translational modifications by a variety of enzymes. Neuropeptides are packaged into and eventually released from the large dense-core vesicles in a regulated secretory pathway. Neuropeptidomics is inherently challenging because neuropeptides are spatially, temporally and chemically heterogeneous in the brain. In addition, the unconventional peptide precursor cleavages and various post-translational modifications require the unbiased measurements of actual peptide forms. Mass spectrometry (MS), coupled with optimized liquid chromatography separation and aided by appropriate sample preparation, has been successfully applied for peptide identification and quantitation. Neuropeptide production regulated by DIMM-dependent genetic mechanism and post-translational processing through alpha-amidation has been studied with different MS-based approaches such as matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) MS and electrospray ionization ion-trap (ESI-IT) MS. DIMM, a transcription factor in Drosophila, appears able to make non-peptidergic cells into peptidergic neurons. Qualitative MS approaches allowed us to observe the fully processed ectopic peptide MII in Drosophila non-peptidergic cells only when DIMM was present. Quantitative MS approaches, together with molecular biological methods, revealed DIMM enabled this complete and efficient ectopic peptide processing through regulating gene CG13248. In addition, an important post-translational modification, peptide alpha-amidation, was studied with mice of different genotypes on different copper diets. The results on the peptide level changes of mature peptides and their immature intermediate forms facilitate our understanding on peptide amidation reaction and its dedicated enzyme peptidylglycine alpha-amidating monooxygenase (PAM). Neurons and glia are two main types of cells in the central nervous system. Analogous to neuropeptidome, the knowledge of the gliopeptidome would aid our understanding of the interaction between glia and neurons. Therefore, the peptides in and regulatedly released from glia were examined. Peptides derived from proteins or putatively from neuropeptide precursors were identified with liquid chromatography coupled to mass spectrometry (LC-MS). Future studies would focus on the confirmation of the putatively identified peptides and the functions of identified peptides in glia. This dissertation demonstrates MS-based approaches are useful tools for probing peptides qualitatively and quantitatively, which would greatly expand our understanding of cell-to-cell communication in the nervous system.

Download or read book Protein and Peptide Analysis by Mass Spectrometry written by J. R. Chapman and published by Springer Science & Business Media. This book was released on 1996-08-19 with total page 368 pages. Available in PDF, EPUB and Kindle. Book excerpt: Leading practitioners authoritatively describe the newest and most effective spectrometric techniques for the analysis of proteins and peptides. The areas covered range from the elucidation of primary and secondary protein structure and the rapid identification of proteins using database techniques to methods for sequencing, as well as methods for the quantitative determination of peptides. Other chapters provide detailed information on the analysis of glycoproteins and glycopeptides and on the use of mass spectrometry to probe the interactions of proteins, both covalent and noncovalent.

Download or read book Mass Spectrometric Application in Neuroscience written by Jingxin Wang and published by . This book was released on 2016 with total page 430 pages. Available in PDF, EPUB and Kindle. Book excerpt: Mass spectrometry based comparative proteomics and peptidomics for neuroscience application have been reported in this thesis using different animal and human models. These work are enabled by improved methods that have been developed for neuropeptide characterization and shotgun proteomics. Chapter 1 presents an overview of the background information of each project described in this thesis. In Chapter 2 we employed a combination of cryostat dissection, heat stabilization, neuropeptide extraction and label-free quantitative neuropeptidomics via a liquid chromatography-high resolution mass spectrometry platform to monitor the feeding-induced changes in neuropeptide expression levels within the nucleus accumbens (Acb). We showed feeding caused the expression level changes of ProSAAS. We further performed a microinjection using ProSAAS neuropeptides to the rat Acb. We showed that at high concentration, big LEN significantly increased rats' food and water intake. In addition, both big LEN and PEN at high concentration altered locomotion and rearing. Chapter 3 details my work in Alzheimer's disease (AD) biomarker discovery. By optimized molecular weight cutoff filtration and in house-constructed database, 645 peptides were identified. Interestingly, 42 proSAAS peptides discovered were exclusively cleaved either from N- or C-termini of the proSAAS protein precursor. Glycoproteins were enriched by lectin affinity chromatography, enabling identification of 795 glycoproteins. One way ANOVA was applied for statistical analysis, a total 15 proteins were found differentially expressed among the three groups. The dynamic changes of transthyretin that exhibited initial increase in MCI followed by decrease in AD were reported for the first time. In Chapter 4, a comparative study of membrane protein in mouse hippocampi was conducted by shotgun proteomics. In this study, AT-1 Tg (acetyltransferase) mouse model that selectively overexpresses human AT-1 in neurons. We found a large set of differentially expressed proteins that are involved in neuronal migration and adhesion, neurite outgrowth, transport of synaptic vesicles, assembly of synaptic connections, and regulation of synaptic activity. Chapters 5 and 6 reported on our method development for neuropeptide characterization and detection. Conclusions and research summaries are presented in Chapter 7. Collectively, this dissertation develops several improved mass spectrometric methods for neuropeptide analysis and showcases several important applications in neuroscience.

Download or read book Mammalian Neuroproteomics and Neuropeptidomics written by James A. Dowell and published by . This book was released on 2008 with total page 168 pages. Available in PDF, EPUB and Kindle. Book excerpt:

Download or read book Targeted Biomarker Quantitation by LC MS written by Naidong Weng and published by John Wiley & Sons. This book was released on 2017-07-31 with total page 397 pages. Available in PDF, EPUB and Kindle. Book excerpt: The first book to offer a blueprint for overcoming the challenges to successfully quantifying biomarkers in living organisms The demand among scientists and clinicians for targeted quantitation experiments has experienced explosive growth in recent years. While there are a few books dedicated to bioanalysis and biomarkers in general, until now there were none devoted exclusively to addressing critical issues surrounding this area of intense research. Target Biomarker Quantitation by LC-MS provides a detailed blueprint for quantifying biomarkers in biological systems. It uses numerous real-world cases to exemplify key concepts, all of which were carefully selected and presented so as to allow the concepts they embody to be easily expanded to future applications, including new biomarker development. Target Biomarker Quantitation by LC-MS primarily focuses on the assay establishment for biomarker quantitation—a critical issue rarely treated in depth. It offers comprehensive coverage of three core areas of biomarker assay establishment: the relationship between the measured biomarkers and their intended usage; contemporary regulatory requirements for biomarker assays (a thorough understanding of which is essential to producing a successful and defendable submission); and the technical challenges of analyzing biomarkers produced inside a living organism or cell. Covers the theory of and applications for state-of-the-art mass spectrometry and chromatography and their applications in biomarker analysis Features real-life examples illustrating the challenges involved in target biomarker quantitation and the innovative approaches which have been used to overcome those challenges Addresses potential obstacles to obtain effective biomarker level and data interpretation, such as specificity establishment and sample collection Outlines a tiered approach and fit-for-purpose assay protocol for target biomarker quantitation Highlights the current state of the biomarker regulatory environment and protocol standards Target Biomarker Quantitation by LC-MS is a valuable resource for bioanalytical scientists, drug metabolism and pharmacokinetics scientists, clinical scientists, analytical chemists, and others for whom biomarker quantitation is an important tool of the trade. It also functions as an excellent text for graduate courses in pharmaceutical, biochemistry and chemistry.

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 Neuropeptides and Other Bioactive Peptides written by Lloyd D. Fricker and published by Morgan & Claypool Publishers. This book was released on 2012 with total page 123 pages. Available in PDF, EPUB and Kindle. Book excerpt: Neuropeptides and peptide hormones represent the largest class of chemical messengers that transmit information from one cell to another. In this review, several decades of research on peptides in cell-cell signaling are summarized, with a focus on neuropeptide discovery, biosynthesis, and function. In addition to covering the well-studied aspects of neuropeptides, emerging concepts are discussed, including classical versus non-classical neuropeptides and direct versus indirect neuropeptides. Other potential functions for peptides in intercellular and intracellular signaling are also discussed. Table of Contents: Overview of Neuropeptides / Neuropeptide Discovery / Neuropeptide Biosynthesis / Neuropeptides After Secretion: Receptors and Peptidases / Representative Neuropeptides / Concluding Remarks and Future Directions / References / Author Biography