Chemical Tools For The Study Of Lipids And Biology
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Chemical Tools For The Study Of Lipids And Biology
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Author : Andrew Rudd
language : en
Publisher:
Release Date : 2019
Chemical Tools For The Study Of Lipids And Biology written by Andrew Rudd and has been published by this book supported file pdf, txt, epub, kindle and other format this book has been release on 2019 with categories.
This dissertation will first explore methods to selectively direct proteins to phospholipid membranes. The natural mechanisms that tether proteins to membranes are typically complex, requiring multiple steps and accessory components. It would be advantageous to develop simplified methods to direct proteins of interest to phospholipid membranes in a single step. We have developed a modular method for membrane localization of proteins by using chemically modified phospholipid anchors capable of covalent attachment to O6-methylguanine DNA methyltransferase (SNAP-tag) fusion proteins. We demonstrate that photocaged lipid precursors enable light-triggered spatial and temporal control over protein localization. The anchoring system is compatible with cell-free expression, allowing for genetic targeting of proteins to lipid membranes of giant unilamellar vesicles. This technique can be used to control membrane curvature effects, similar to what has been previously observed with certain membrane-bound proteins. This work addresses a current need in synthetic biology for simplified and robust methods to control membrane localization of expressed proteins and shows promise as a general tool for protein targeting to lipid vesicles and cellular membranes. In the second investigation, this dissertation will discuss a new method for the nonenzymatic remodeling of lipid membranes. Cell membranes have a vast repertoire of phospholipid species whose structures can be dynamically modified by enzymatic remodeling of acyl chains and polar head groups. Lipid remodeling plays important roles in membrane biology and dysregulation can lead to disease. Although there have been tremendous advances in creating artificial membranes to model the properties of native membranes, a major obstacle has been developing straightforward methods to mimic lipid membrane remodeling. Stable liposomes are typically kinetically trapped and are not prone to exchanging diacylphospholipids. An approach relies on transthioesterification/acyl shift reactions that occur spontaneously and reversibly between tertiary amides and thioesters. We demonstrate exchange and remodeling of both lipid acyl chains and head groups. Using our synthetic model system we demonstrate the ability of spontaneous phospholipid remodeling to trigger changes in vesicle spatial organization, composition, and morphology as well as recruit proteins that can affect vesicle curvature. Membranes capable of chemically exchanging lipid fragments could be used to help further understand the specific roles of lipid structure remodeling in biological membranes. In the third section of this dissertation, a novel approach for the in situ synthesis of natural lipids in living cells will be presented. Mammalian cells synthesize thousands of distinct lipids, yet the function of many of these lipid species is unknown. Ceramides, a class of sphingolipid, are implicated in several cell-signaling pathways but poor cell permeability and lack of selectivity in endogenous synthesis pathways have hampered direct study of their effects. Here we report a strategy that overcomes the inherent biological limitations of ceramide delivery by chemoselectively ligating lipid precursors in vivo to yield natural ceramides in a traceless manner. Using this method, we uncovered the apoptotic effects of several ceramide species and observed differences in their apoptotic activity based on acyl-chain saturation. Additionally, we demonstrate spatiotemporally controlled ceramide synthesis in live cells through photoinitiated lipid ligation. Our in situ lipid ligation approach addresses the long-standing problem of lipid-specific delivery and enables the direct study of unique ceramide species in live cells. Finally, this dissertation will discuss the development of small molecules for the depalmitoylation of proteins in vivo. Post-translational S-palmitoylation plays a central role in protein localization, trafficking, stability, aggregation, and cell signaling. Dysregulation of palmitoylation pathways in cells can alter protein function and is the cause of several diseases. Considering the biological and clinical importance of S-palmitoylation, tools for direct, in vivo modulation of this lipid modification would be extremely valuable. Here, we describe a method for the cleavage of native S-palmitoyl groups from proteins in living cells. Using a cell permeable, cysteine-functionalized amphiphile, we demonstrate the direct depalmitoylation of cellular proteins. We show that amphiphile-mediated depalmitoylation (AMD) can effectively cleave S-palmitoyl groups from the native GTPase HRas and successfully depalmitoylate mislocalized proteins in an infantile neuronal ceroid lipofuscinosis (INCL) disease model. AMD enables direct and facile depalmitoylation of proteins in live cells and has potential therapeutic applications for diseases such as INCL, where native protein thioesterase activity is deficient.
Fatty Acid And Lipid Chemistry
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Author : F.D. Gunstone
language : en
Publisher: Springer
Release Date : 2012-12-06
Fatty Acid And Lipid Chemistry written by F.D. Gunstone and has been published by Springer this book supported file pdf, txt, epub, kindle and other format this book has been release on 2012-12-06 with Science categories.
This book has a pedigree. It has developed from earlier publications by the author and from his experience over 50 years in reading, writing, thinking, and working with lipids and fatty acids. The earlier publications are: (i) An Introduction to the Chemistry of Fats and Fatty Acids, Chapman and Hall, 1958. (ii) An Introduction to the Chemistry and Biochemistry of Fatty Acids and their Glycerides, Chapman and Hall, 1967. (iii) Lipids in Foods: Chemistry, Biochemistry, and Technology (with F. A. Norris), Pergamon Press, 1983. (iv) The Lipid Handbook (with J. L. Harwood and F. B. Padley), Chapman and Hall, first edition 1986, second edition 1994. (v) A Lipid Glossary (with B. G. Herslof), The Oily Press, Dundee, 1992. (vi) Lecture notes for a course on Fatty Acids and Lipids designed for those entering the oil and fat industry and given on over 20 occasions since 1977. The book is dedicated to the next generation of lipid scientists. The study of lipids now involves many disciplines, all of which require a basic knowledge of the chemical nature and properties of these molecules, which is what this book is about. It is written particularly for those who, with some knowledge of chemistry or biochemistry, need to know more about the nature of lipids and fatty acids.
Current Protocols In Chemical Biology
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Author :
language : en
Publisher:
Release Date :
Current Protocols In Chemical Biology written by and has been published by this book supported file pdf, txt, epub, kindle and other format this book has been release on with Biochemistry categories.
Current Protocols in Chemical Biology provides validated protocols and enlightening overviews for researchers requiring specialized chemical techniques and tools for their studies of biology and drug design. Current Protocols in Chemical Biology describes these chemical tools, including small-molecule design, synthesis, derivatization, handling, and detection. Advances in laboratory automation, robotics, and medicinal chemistry will be described as applied to high-throughput screening (HTS) methods. Also included will be methods for modification of proteins, nucleic acids, carbohydrates and lipids for their use as tools in the study of particular biological systems. Investigators in the fields of medicinal chemistry, combinatorial chemistry, proteomics, pharmacology, biochemistry, glycobiology, chemical genetics, molecular biology, cell biology, and cytometry will find relevant, validated, state-of-the-art methods in Current Protocols in Chemical Biology.
Bioanalytics
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Author : Friedrich Lottspeich
language : en
Publisher: John Wiley & Sons
Release Date : 2018-05-29
Bioanalytics written by Friedrich Lottspeich and has been published by John Wiley & Sons this book supported file pdf, txt, epub, kindle and other format this book has been release on 2018-05-29 with Science categories.
Analytical methods are the essential enabling tools of the modern biosciences. This book presents a comprehensive introduction into these analytical methods, including their physical and chemical backgrounds, as well as a discussion of the strengths and weakness of each method. It covers all major techniques for the determination and experimental analysis of biological macromolecules, including proteins, carbohydrates, lipids and nucleic acids. The presentation includes frequent cross-references in order to highlight the many connections between different techniques. The book provides a bird's eye view of the entire subject and enables the reader to select the most appropriate method for any given bioanalytical challenge. This makes the book a handy resource for students and researchers in setting up and evaluating experimental research. The depth of the analysis and the comprehensive nature of the coverage mean that there is also a great deal of new material, even for experienced experimentalists. The following techniques are covered in detail: - Purification and determination of proteins - Measuring enzymatic activity - Microcalorimetry - Immunoassays, affinity chromatography and other immunological methods - Cross-linking, cleavage, and chemical modification of proteins - Light microscopy, electron microscopy and atomic force microscopy - Chromatographic and electrophoretic techniques - Protein sequence and composition analysis - Mass spectrometry methods - Measuring protein-protein interactions - Biosensors - NMR and EPR of biomolecules - Electron microscopy and X-ray structure analysis - Carbohydrate and lipid analysis - Analysis of posttranslational modifications - Isolation and determination of nucleic acids - DNA hybridization techniques - Polymerase chain reaction techniques - Protein sequence and composition analysis - DNA sequence and epigenetic modification analysis - Analysis of protein-nucleic acid interactions - Analysis of sequence data - Proteomics, metabolomics, peptidomics and toponomics - Chemical biology
Chemical And Synthetic Approaches In Membrane Biology
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Author : Arun K. Shukla
language : en
Publisher: Humana Press
Release Date : 2017-06-12
Chemical And Synthetic Approaches In Membrane Biology written by Arun K. Shukla and has been published by Humana Press this book supported file pdf, txt, epub, kindle and other format this book has been release on 2017-06-12 with Science categories.
This practical volume brings together a broad range of topics related to membrane biology research with particular emphasis on novel approaches, technology platforms, and emerging tools in this area of study. Beginning with chapters pertaining to artificial or designer membrane mimetics that can be utilized for in vitro studies of membrane-lipid interactions, it then continues by covering the approaches and methodologies to directly investigate membrane protein structure, localization, and dynamics. Written for the Springer Protocols Handbooks series, this collection focuses on in-depth, hands-on protocols from experts in the field. Authoritative and cutting-edge, Chemical and Synthetic Approaches in Membrane Biology serves as an ideal guide to researchers investigating the vital properties of cellular membranes.
Polar Lipids
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Author : Moghis U. Ahmad
language : en
Publisher: Elsevier
Release Date : 2015-08-13
Polar Lipids written by Moghis U. Ahmad and has been published by Elsevier this book supported file pdf, txt, epub, kindle and other format this book has been release on 2015-08-13 with Science categories.
Polar Lipids is a valuable reference resource providing thorough and comprehensive coverage of different types of polar lipids known to lipid science and industry today. This book covers important applications and utilization of polar lipids, either in the area of food and nutrition, or health and disease. Each chapter covers chemistry and chemical synthesis, biosynthesis and biological effects, functional and nutritional properties, applications, processing technologies, and future trends of a variety of polar lipids—including glycolipids, ether lipids, phenol lipids, serine phospholipids, omega-3 phospholipids, rice lecithin, palm lecithin, sunflower lecithin, sugar- and protein-based lipids, lysophospholipids,and more. Presents new and relatively unexplored polar lipids for researchers to consider to use in food and health applications Includes details on the chemistry and chemical synthesis, biosynthesis and biological effects, functional and nutritional properties, applications, and future trends of a variety of polar lipids Presents the latest analytical techniques for use in polar lipids research, including NMR and Supercritical Fluid Chromatography/Mass Spectrometry
Chemical Tools To Study Lipid Signaling
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Author :
language : en
Publisher:
Release Date : 2020
Chemical Tools To Study Lipid Signaling written by and has been published by this book supported file pdf, txt, epub, kindle and other format this book has been release on 2020 with categories.
Lipid Biochemistry
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Author : J.L. Harwood
language : en
Publisher: Springer Science & Business Media
Release Date : 2013-04-17
Lipid Biochemistry written by J.L. Harwood and has been published by Springer Science & Business Media this book supported file pdf, txt, epub, kindle and other format this book has been release on 2013-04-17 with Science categories.
Lipids can usually be extracted easily from tissues by making use of their hydrophobic characteristics. However, such extractions yield a complex mixture of different lipid classes which have to be purified further for quantitative analysis. Moreover, the crude lipid extract will be contami nated by other hydrophobic molecules, e.g. by intrinsic membrane proteins. Of the various types of separation processes, thin layer and column chromatography are most useful for intact lipids. High performance liquid chromatography (HPLC) is also rapidly becoming more popular, especially for the fractionation of molecular species of a given lipid class. The most powerful tool for quantitation of the majority of lipids is gas liquid chromatography (GLC). The method is very sensitive and, if adapted with capillary columns, can provide information with regard to such subtle features as the position or configuration of substitutions along acyl chains. By coupling GLC or HPLC to a radioactivity detector, then the techniques are also very useful for metabolic measurements. Although research laboratories use generally sophisticated analytical methods such as GLC to analyse and quantify lipid samples, chemical derivatie:ations are often used in hospitals. For these methods, the lipid samples are derivatized to yield a product which can be measured simply and accurately-usually by colour. Thus, total triacylglycerol, cholesterol or phospholipid-phosphorus can be quantitated conveniently without bothering with the extra information of molecular species, etc. which might be determined by more thorough analyses. REFERENCES Christie, w.w. (1982) Lipid Analysis, 2nd edn, Pergamon Press, Oxford.
Plant Lipids
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Author : Denis J. Murphy
language : en
Publisher: CRC Press
Release Date : 2020-02-03
Plant Lipids written by Denis J. Murphy and has been published by CRC Press this book supported file pdf, txt, epub, kindle and other format this book has been release on 2020-02-03 with Science categories.
New research tools have revealed many surprising aspects of the dynamic nature of lipids and their participation in processes such as recognition, intra- and inter-cellular signalling, deterrence and defense against pathogens, membrane trafficking and protein function. This is in addition to new information on the more established roles of plant lipids as structural components of membranes and as long-term storage products. Plant lipids are also increasingly being seen as sources of a new generation of environmentally friendly, biodegradable, and renewable industrial products, including biopolymers and high-grade lubricants. Plant Lipids: Biology, Utilisation and Manipulation provides a broad overview of plant lipid research and its many applications. Linking various disciplines, the editor brings together researchers from major international laboratories to review the history and current state of progress in this quickly evolving field. The text starts by providing a fascinating historical perspective on the study of plant lipids, from its inception as a branch of alchemy in the seventeenth century to the current post-genomic era. It then offers a detailed discussion on the formation, modification and utilization of fatty acids. This is followed by an exploration of the major classes of macromolecular structures formed by plant lipids, including bilayer membranes and storage bodies. From there, the contributors consider other types of macromolecular lipid assemblies in plants, examining proteins and the key plant lipid structure - the cuticle. The final chapters look at diverse classes of plant lipids that are linked to various aspects of signaling. This text provides an excellent resource for researchers and professionals in plant biochemistry, molecular biology, biotechnology and genetics, in both the academic and industrial sectors. It also meets the needs of students looking for a comprehensive introduction to this field, as well as direction for fut
Mass Spectrometry For Lipidomics
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Author : Michal Holcapek
language : en
Publisher: John Wiley & Sons
Release Date : 2023-03-07
Mass Spectrometry For Lipidomics written by Michal Holcapek and has been published by John Wiley & Sons this book supported file pdf, txt, epub, kindle and other format this book has been release on 2023-03-07 with Science categories.
Mass Spectrometry for Lipidomics All-in-one guide to successful lipidomic analysis, combining the latest advances and best practices from academia, industry, and clinical research Mass Spectrometry for Lipidomics presents a systematic overview of lipidomic analysis, covering established standards of lipid analysis, available technology, and key lipid classes, as well as applications in basic research, medicine, pharma, and the food industry. Through connecting recent technological advances with key application areas, this unique guide bridges the gap between academia and industry by translating the vast body of knowledge that has been gained in the past decade into much-needed practical advice for novices as well as routine users. Edited by the president and vice-president of the International Lipidomics Society with contributions from the top experts in lipid analysis, Mass Spectrometry for Lipidomics covers a wide range of key topics, including: Aspects of sample preparation, separation methods, different mass spectrometry modes, as well as identification and quantitation, including the use of bioinformatics tools for data analysis Identification, quantitation and profiling of lipids in different types of biological samples Analytical approaches for all major classes of biological lipids, from fatty acids to phospholipids to sterols Novel applications in biological research, clinical diagnostics, as well as food and crop science For analytical chemists, biochemists, clinical chemists, and analytical laboratories and hospitals, Mass Spectrometry for Lipidomics presents a comprehensive and authoritative overview of the subject, with unmatched expertise from practicing professionals actively involved in the latest research.