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Author: Jianyong Wu Publisher: ISBN: Category : Oxidative stress Languages : en Pages : 434
Book Description
The adduction of proteins and other biomolecules by electrophilic lipid peroxidation products such as 4-hydroxy-2-nonenal (HNE), 4-oxo-2-nonenal (ONE), malondialdehyde (MDA) or acrolein (ACR) is thought to be an initiating and/or propagating factor in the pathophysiology of several diseases such as atherosclerosis, diabetes, Alzheimer's, Parkinson's and other age-related disorders. The identification of protein sites modified by oxylipids is of key relevance for advancing our understanding how oxidative damage affects structure and function of proteins. Here, the use of MALDI tandem mass spectrometry with high energy collision-induced dissociation (CID) on a TOF/TOF instrument for sequencing oxylipid-peptide conjugates was systematically studied. Three synthesized model peptides containing one nucleophilic residue (i.e. Cys, His or Lys) were reacted with MDA, HNE, ONE and ACR. MALDI-MS analysis and MS/MS analysis were performed to confirm the adduct type and the modification sites. Michael adducts and Schiff bases were the predominant products under pH 7.4 within 2 hours. All MS/MS spectra of Michael adducts show the neutral loss of the oxylipid moiety ions. MS/MS spectra of Cys-containing peptide oxylipid conjugates exhibit additional characteristic neutral loss of HS-oxylipid moiety ions. MS/MS spectra of His-containing peptide oxylipid conjugates show characteristic oxylipid-containing His immonium ions. Spectra of Lys-containing peptide oxylipid conjugates (Schiff base) also show oxylipid-containing Lys immonium ions. However, there is no neutral loss of the oxylipid moiety ion for these Schiff bases. Determining the extent or relative amounts of the oxidative damage in cells could provide valuable insights into the molecular mechanisms of the diseases caused by oxidative stress. Relative quantitation of oxylipid-modified proteins in biological samples is a challenging problem because of the complexity and extreme dynamic range that characterize these samples. In this study, the reagents, N'-aminooxymethylcarbonylhydrazino-D-biotin (ARP) and iodoacetyl-PEO2- biotin (IPB), were used to enrich acrolein-modified Cys-containing peptides and the corresponding unmodified ones from subsarcolemmal mitochondria (SSM). The ratios between them were determined by nanoLC-SRM analysis. Model Cys-containing peptides labeled with ARP-acrolein and IPB were employed to demonstrate this method. Seven acrolein-modifed Cys-containing peptides from five mitochondrial proteins were quantified. The ratios for those seven peptides from the CCl4-treated rats are higher than the control ones indicating that the ratios of acrolein-modified peptides to unmodified ones are potential markers of oxidative stress in vivo. Age-dependent changes of protein carbonyls were investigated in subsarcolemmal mitochondria by using LC-SRM analysis of distinct ACR-modified Cys-containing peptides. Immunochemical analysis using an anti-ACR monoclonal antibody supported an increase of proteins modified by acrolein with age. However, total protein carbonyls measurement using ARP in Western blot analysis did not conform to this change suggesting that age-related changes in protein carbonyls are complex and would benefit from more specific measurement protocols.
Author: Jianyong Wu Publisher: ISBN: Category : Oxidative stress Languages : en Pages : 434
Book Description
The adduction of proteins and other biomolecules by electrophilic lipid peroxidation products such as 4-hydroxy-2-nonenal (HNE), 4-oxo-2-nonenal (ONE), malondialdehyde (MDA) or acrolein (ACR) is thought to be an initiating and/or propagating factor in the pathophysiology of several diseases such as atherosclerosis, diabetes, Alzheimer's, Parkinson's and other age-related disorders. The identification of protein sites modified by oxylipids is of key relevance for advancing our understanding how oxidative damage affects structure and function of proteins. Here, the use of MALDI tandem mass spectrometry with high energy collision-induced dissociation (CID) on a TOF/TOF instrument for sequencing oxylipid-peptide conjugates was systematically studied. Three synthesized model peptides containing one nucleophilic residue (i.e. Cys, His or Lys) were reacted with MDA, HNE, ONE and ACR. MALDI-MS analysis and MS/MS analysis were performed to confirm the adduct type and the modification sites. Michael adducts and Schiff bases were the predominant products under pH 7.4 within 2 hours. All MS/MS spectra of Michael adducts show the neutral loss of the oxylipid moiety ions. MS/MS spectra of Cys-containing peptide oxylipid conjugates exhibit additional characteristic neutral loss of HS-oxylipid moiety ions. MS/MS spectra of His-containing peptide oxylipid conjugates show characteristic oxylipid-containing His immonium ions. Spectra of Lys-containing peptide oxylipid conjugates (Schiff base) also show oxylipid-containing Lys immonium ions. However, there is no neutral loss of the oxylipid moiety ion for these Schiff bases. Determining the extent or relative amounts of the oxidative damage in cells could provide valuable insights into the molecular mechanisms of the diseases caused by oxidative stress. Relative quantitation of oxylipid-modified proteins in biological samples is a challenging problem because of the complexity and extreme dynamic range that characterize these samples. In this study, the reagents, N'-aminooxymethylcarbonylhydrazino-D-biotin (ARP) and iodoacetyl-PEO2- biotin (IPB), were used to enrich acrolein-modified Cys-containing peptides and the corresponding unmodified ones from subsarcolemmal mitochondria (SSM). The ratios between them were determined by nanoLC-SRM analysis. Model Cys-containing peptides labeled with ARP-acrolein and IPB were employed to demonstrate this method. Seven acrolein-modifed Cys-containing peptides from five mitochondrial proteins were quantified. The ratios for those seven peptides from the CCl4-treated rats are higher than the control ones indicating that the ratios of acrolein-modified peptides to unmodified ones are potential markers of oxidative stress in vivo. Age-dependent changes of protein carbonyls were investigated in subsarcolemmal mitochondria by using LC-SRM analysis of distinct ACR-modified Cys-containing peptides. Immunochemical analysis using an anti-ACR monoclonal antibody supported an increase of proteins modified by acrolein with age. However, total protein carbonyls measurement using ARP in Western blot analysis did not conform to this change suggesting that age-related changes in protein carbonyls are complex and would benefit from more specific measurement protocols.
Author: Juan D. Chavez Publisher: ISBN: Category : Active oxygen Languages : en Pages : 702
Book Description
Oxidative stress is recognized as an important underlying factor in the pathogenesis of many degenerative diseases as well as normal senescence. The free radicals, reactive oxygen species (ROS) and electrophiles produced during oxidative stress are capable of modifying nucleic acids, lipids and proteins. There are a variety of oxidative modifications that occur to proteins including: cleavage of the protein backbone, direct oxidation of amino acid side chains by ROS, and adduction by electrophilic species such as lipid peroxidation products. Many of these oxidative modifications result in the introduction of carbonyl groups into the proteins. Protein carbonylation levels are commonly used as a biomarker to assess the degree of oxidative damage to a system. However the most commonly employed methods for measuring oxidative modifications to proteins, typically fail to provide any information about the identity of the modified protein, site of modification, or the chemical nature of the modification. In the present study we develop an analytical technique based on affinity labeling with N'-aminooxymethylcarbonylhydrazino-D-biotin (aldehyde reactive probe, ARP), along with mass spectrometric analysis which allows for the full characterization of protein carbonylation modifications. The ability of the ARP method was first demonstrated for the case of oxylipid peptide and protein conjugates formed by Michael addition-type conjugation reactions with [alpha,beta]- unsaturated aldehydic lipid peroxidation products with nucleophilic amino acid residue side chains. ARP was used to label a 4-hydroxy-2-nonenal (HNE) modified cysteine containing model peptide, and HNE modified E. coli thioredoxin, which were characterized using ESI-MS/MS and MALDI-MS/MS. ARP was also used to label the oxidative modifications alpha-aminoadipic semialdehyde (AAS) and gamma-glutamic semialdehyde (GGS), formed during the metal catalyzed oxidation of GAPDH. After demonstrating the utility of the technique on model systems, it was then applied to complex biological systems. In one case, subsarcolemmal mitochondria (SSM) isolated from rat cardiac tissue. Mitochondria are well known to be a major source of ROS within the cell. They are therefore important mediators of oxidative stress, as well as regulators of cell death. We were able to identify 39 unique sites on 27 mitochondrial proteins which were modified by six different [alpha,beta]-unsaturated aldehydes, including acrolein, [beta]-hydroxyacrolein, crotonaldehyde, 4-hydroxy-2-hexenal, 4-hydroxy-2-nonenal and 4-oxo-2- nonenal. Additionally we identified nine Lys residues on four mitochondrial proteins that were oxidized to AAS and subsequently labeled with ARP. The proteins identified with oxidative modifications include members of the mitochondrial electron transport chain, TCA cycle, membrane transport, lipid metabolism, and other important mitochondrial enzymes. The ARP technique was also applied to identify protein targets of 4-hyroxy-2- nonenal in human monocytic THP-1 cells that were exogenously exposed to HNE. It was shown previously that exposure of THP-1 cells to HNE resulted in apoptosis, necrosis and protein carbonylation. We applied a multi-pronged proteomic approach involving electrophoretic, immunoblotting and mass spectrometric analysis to unequivocally identify eighteen sites of HNE modification on sixteen proteins. It was also demonstrated in this study that pretreatment of THP-1 cells with ascorbic acid resulted in decreased levels of HNE-protein conjugate formation.
Author: Lawrence J. Berliner Publisher: Springer Nature ISBN: 303047318X Category : Science Languages : en Pages : 237
Book Description
This book describes the methods of analysis and determination of oxidants and oxidative stress in biological systems. Reviews and protocols on select methods of analysis of ROS, RNS, oxygen, redox status, and oxidative stress in biological systems are described in detail. It is an essential resource for both novices and experts in the field of oxidant and oxidative stress biology.
Author: Publisher: Academic Press ISBN: 0123813468 Category : Science Languages : en Pages : 411
Book Description
Thiol Redox Transitions in Cell Signaling, Part A, along with its companion (volume 475), presents methods and protocols dealing with thiol oxidation-reduction reactions and their implications as they relate to cell signaling. This first installment of Cadenas and Packer's two-volume treatment specifically deals with glutathionylation and dethiolation, and peroxide removal by peroxiredoxins/thioredoxins and glutathione peroxidases. The critically acclaimed laboratory standard for 40 years, Methods in Enzymology is one of the most highly respected publications in the field of biochemistry. Since 1955, each volume has been eagerly awaited, frequently consulted, and praised by researchers and reviewers alike. Over 450 volumes have been published to date, and much of the material is relevant even today--truly an essential publication for researchers in all fields of life sciences. Along with companion volume, provides a full overview of techniques necessary to the study of thiol redox in relation to cell signaling Gathers tried and tested techniques from global labs, offering both new and tried-and-true methods Relevant background and reference information given for procedures can be used as a guide to developing protocols in a number of disciplines
Author: Kazuwa Nakao Publisher: Springer ISBN: 4431556516 Category : Science Languages : en Pages : 330
Book Description
This book is devoted to innovative medicine, comprising the proceedings of the Uehara Memorial Foundation Symposium 2014. It remains extremely rare for the findings of basic research to be developed into clinical applications, and it takes a long time for the process to be achieved. The task of advancing the development of basic research into clinical reality lies with translational science, yet the field seems to struggle to find a way to move forward. To create innovative medical technology, many steps need to be taken: development and analysis of optimal animal models of human diseases, elucidation of genomic and epidemiological data, and establishment of “proof of concept”. There is also considerable demand for progress in drug research, new surgical procedures, and new clinical devices and equipment. While the original research target may be rare diseases, it is also important to apply those findings more broadly to common diseases. The book covers a wide range of topics and is organized into three complementary parts. The first part is basic research for innovative medicine, the second is translational research for innovative medicine, and the third is new technology for innovative medicine. This book helps to understand innovative medicine and to make progress in its realization.
Author: Isabella Dalle-Donne Publisher: John Wiley & Sons ISBN: 0471973114 Category : Science Languages : en Pages : 978
Book Description
Methodology and applications of redox proteomics The relatively new and rapidly changing field of redox proteomics has the potential to revolutionize how we diagnose disease, assess risks, determine prognoses, and target therapeutic strategies for people with inflammatory and aging-associated diseases. This collection brings together, in one comprehensive volume, a broad array of information and insights into normal and altered physiology, molecular mechanisms of disease states, and new applications of the rapidly evolving techniques of proteomics. Written by some of the finest investigators in this area, Redox Proteomics: From Protein Modifications to Cellular Dysfunction and Diseases examines the key topics of redox proteomics and redox control of cellular function, including: * The role of oxidized proteins in various disorders * Pioneering studies on the development of redox proteomics * Analytical methodologies for identification and structural characterization of proteins affected by oxidative/nitrosative modifications * The response and regulation of protein oxidation in different cell types * The pathological implications of protein oxidation for conditions, including asthma, cardiovascular disease, diabetes, preeclampsia, and Alzheimer's disease Distinguished by its in-depth discussions, balanced methodological approach, and emphasis on medical applications and diagnosis development, Redox Proteomics is a rich resource for all professionals with an interest in proteomics, cellular physiology and its alterations in disease states, and related fields.
Author: United States. Public Health Service. Office of the Surgeon General Publisher: ISBN: Category : Government publications Languages : en Pages : 728
Book Description
This report considers the biological and behavioral mechanisms that may underlie the pathogenicity of tobacco smoke. Many Surgeon General's reports have considered research findings on mechanisms in assessing the biological plausibility of associations observed in epidemiologic studies. Mechanisms of disease are important because they may provide plausibility, which is one of the guideline criteria for assessing evidence on causation. This report specifically reviews the evidence on the potential mechanisms by which smoking causes diseases and considers whether a mechanism is likely to be operative in the production of human disease by tobacco smoke. This evidence is relevant to understanding how smoking causes disease, to identifying those who may be particularly susceptible, and to assessing the potential risks of tobacco products.
Author: Mike S. Lee Publisher: John Wiley & Sons ISBN: 1118180720 Category : Science Languages : en Pages : 1362
Book Description
Due to its enormous sensitivity and ease of use, mass spectrometry has grown into the analytical tool of choice in most industries and areas of research. This unique reference provides an extensive library of methods used in mass spectrometry, covering applications of mass spectrometry in fields as diverse as drug discovery, environmental science, forensic science, clinical analysis, polymers, oil composition, doping, cellular research, semiconductor, ceramics, metals and alloys, and homeland security. The book provides the reader with a protocol for the technique described (including sampling methods) and explains why to use a particular method and not others. Essential for MS specialists working in industrial, environmental, and clinical fields.
Author: Victor R Preedy Publisher: Royal Society of Chemistry ISBN: 1782622616 Category : Medical Languages : en Pages : 624
Book Description
Carnosine and anserine are two common histidine-derived dipeptides, also known as imidazole dipeptides, found in muscle and brain tissues of animals. Their dietary and nutritional significance lies in their antioxidant properties and a number of investigators are looking at other health benefits of the compounds. This volume surveys the current state of play in these and other areas of interest, including chemistry and biochemistry, immunology and cellular homeostasis. Chapters look in depth at some of the functions and effects in the body particularly with regard to exercise and fatigue, wound healing, cancer treatment and age-related diseases. Providing an up to date, interdisciplinary approach, this book is of great interest to researchers and professionals in chemistry, food science, nutrition, biochemistry, health sciences and sports sciences.
Author: Jianyong Wu
Author: Jianyong Wu
Author: Juan D. Chavez
Author: Lawrence J. Berliner
Author: 
Author: Kazuwa Nakao
Author: Isabella Dalle-Donne
Author: United States. Public Health Service. Office of the Surgeon General
Author: 
Author: Mike S. Lee
Author: Victor R Preedy