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Heterologous Expression Of Membrane Proteins
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Book Synopsis Membrane Protein Protocols by : Barry S. Selinsky
Download or read book Membrane Protein Protocols written by Barry S. Selinsky and published by Springer Science & Business Media. This book was released on 2008-02-03 with total page 330 pages. Available in PDF, EPUB and Kindle. Book excerpt: Knowledge of the three-dimensional structure of a protein is absolutely required for the complete understanding of its function. The spatial orientation of amino acids in the active site of an enzyme demonstrates how substrate specificity is defined, and assists the medicinal chemist in the design of s- cific, tight-binding inhibitors. The shape and contour of a protein surface hints at its interaction with other proteins and with its environment. Structural ana- sis of multiprotein complexes helps to define the role and interaction of each individual component, and can predict the consequences of protein mutation or conditions that promote dissociation and rearrangement of the complex. Determining the three-dimensional structure of a protein requires milligram quantities of pure material. Such quantities are required to refine crystallization conditions for X-ray analysis, or to overcome the sensitivity limitations of NMR spectroscopy. Historically, structural determination of proteins was limited to those expressed naturally in large amounts, or derived from a tissue or cell source inexpensive enough to warrant the use of large quantities of cells. H- ever, with the advent of the techniques of modern gene expression, many p- teins that are constitutively expressed in minute amounts can become accessible to large-scale purification and structural analysis.
Book Synopsis Heterologous Gene Expression in E.coli by : Thomas C. Evans, Jr.
Download or read book Heterologous Gene Expression in E.coli written by Thomas C. Evans, Jr. and published by Humana Press. This book was released on 2011-08-24 with total page 310 pages. Available in PDF, EPUB and Kindle. Book excerpt: Protein expression in a heterologous host is a cornerstone of biomedical research and of the biotechnology industry. Despite the advanced state of protein expression technology improvements are still needed. For example, membrane proteins constitute a significant percentage of the total cellular proteins but as a class are very difficult to overexpress, especially in a heterologous host. The ideal host would have the ability to express any protein, with relevant post-translational modifications, and be as easy to work with as E. coli. In Heterologous Gene Expression in E. coli: Methods and Protocols, expert scientists intimately familiar with the relevant techniques offer chapters that greatly expand the utility of this expression host. The contributions in this detailed volume describe methods, for example, to successfully express proteins in E. coli that would otherwise form aggregates in this host, to add post-translational modifications, to incorporate non-standard amino acid residues or moieties into E. coli expressed proteins, to identify binding partners, and to express membrane proteins. Written in the highly successful Methods in Molecular BiologyTM format, chapters include introductions to their respective subjects, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Practical and cutting-edge, Heterologous Gene Expression in E. coli: Methods and Protocols seeks to familiarize the researcher with the myriad of E. coli expression strains available and move E. coli closer to that ideal of the perfect host.
Book Synopsis Heterologous Gene Expression in E.coli by : Nicola A. Burgess-Brown
Download or read book Heterologous Gene Expression in E.coli written by Nicola A. Burgess-Brown and published by Humana Press. This book was released on 2018-07-20 with total page 429 pages. Available in PDF, EPUB and Kindle. Book excerpt: This detailed volume provides a toolbox for designing constructs, tackling expression and solubility issues, handling membrane proteins and protein complexes, and exploring innovative engineering of E. coli. The topics are largely grouped under four parts: high-throughput cloning, expression screening, and optimization of expression conditions, protein production and solubility enhancement, case studies to produce challenging proteins and specific protein families, as well as applications of E. coli expression. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Heterologous Gene Expression in E. coli: Methods and Protocols serves molecular biologists, biochemists and structural biologists, those in the beginning of their research careers to those in their prime, to give both an historical and modern overview of the methods available to express their genes of interest in this exceptional organism.
Book Synopsis Heterologous Expression of Membrane Proteins by : Isabelle Mus-Veteau
Download or read book Heterologous Expression of Membrane Proteins written by Isabelle Mus-Veteau and published by Springer Nature. This book was released on 2022-06-30 with total page 472 pages. Available in PDF, EPUB and Kindle. Book excerpt: This detailed volume explores protocols for the production of membrane proteins in a panel of heterologous organisms for structural studies. Beginning with techniques using E. coli as a host for the overproduction and purification of membrane proteins, the book continues with chapters covering mammalian membrane protein production in yeast, insect cells, mammalian cells, as well as using virus like particles and acellular systems. Additionally, new detergents and alternatives to detergents allowing membrane protein purification for structural analyses are described. The book closes with a chapter exploring the use of microscale thermophoresis (MST) to evaluate the binding activity of heterologously expressed proteins directly in crude membrane extracts. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and up-to-date, Heterologous Expression of Membrane Proteins: Methods and Protocols, Third Edition serves as an ideal guide for scientists aiming to produce and purify functional recombinant membrane proteins for structural studies.
Book Synopsis The Biochemistry of Archaea (Archaebacteria) by : M. Kates
Download or read book The Biochemistry of Archaea (Archaebacteria) written by M. Kates and published by Elsevier. This book was released on 1993-12-13 with total page 633 pages. Available in PDF, EPUB and Kindle. Book excerpt: In the last 10 years, considerable information has accumulated on the biochemistry of archaea. In this volume, the subject as a whole is treated in a comprehensive manner. The book brings together recent knowledge concerning general metabolism, bioenergetics, molecular biology and genetics, membrane lipid and cell-wall structural chemistry and evolutionary relations, of the three major groups of archaea: the extreme halophiles, the extreme thermophiles, and the methanogens.Subjects included are: the evolutionary relationship of these microorganisms to all other living cells; special metabolic features of archeaea; protein structural chemistry; cell envelopes; molecular biology in archaea including DNA structure and replication, transcription apparatus, translation apparatus, and ribosomal structure; and a final chapter on the molecular genetics of archaea. This comprehensive scope ensures its usefulness to researchers, and stimulates further study in this rapidly developing field.
Book Synopsis Chemical and Synthetic Biology Approaches to Understand Cellular Functions - Part C by :
Download or read book Chemical and Synthetic Biology Approaches to Understand Cellular Functions - Part C written by and published by Academic Press. This book was released on 2020-02-10 with total page 310 pages. Available in PDF, EPUB and Kindle. Book excerpt: Chemical and Synthetic Biology Approaches to Understand Cellular Functions - Part C, Volume 633, the latest release in the Methods in Enzymology series, continues the legacy of this premier serial. This release includes sections on Next generation probes for molecular imaging in cells, Competitive binding assay for biotin and biotin derivatives, based on avidin and biotin-4-fluorescein, Converting avidin to bind ligands other than biotin, especially steroids, Chemoenzymatic Labeling Strategy, Engineered Siderophores, Small molecules to inhibit bacterial population behavior, NMR tube bioreactor, Small molecule controlled RAS activation system, Small molecule regulated Cas9, the Design and application of synthetic receptors, and much more. - Contains the authority of authors who are leaders in their field - Provides a comprehensive source on new methods and research in enzymology
Book Synopsis Production of Membrane Proteins by : Anne Skaja Robinson
Download or read book Production of Membrane Proteins written by Anne Skaja Robinson and published by John Wiley & Sons. This book was released on 2011-06-15 with total page 631 pages. Available in PDF, EPUB and Kindle. Book excerpt: Designed as a research-level guide to current strategies and methods of membrane protein production on the small to intermediate scale, this practice-oriented book provides detailed, step-by-step laboratory protocols as well as an explanation of the principles behind each method, together with a discussion of its relative advantages and disadvantages. Following an introductory section on current challenges in membrane protein production, the book goes on to look at expression systems, emerging methods and approaches, and protein specific considerations. Case studies illustrate how to select or sample the optimal production system for any desired membrane protein, saving both time and money on the laboratory as well as the technical production scale. Unique in its coverage of "difficult" proteins with large membrane-embedded domains, proteins from extremophiles, peripheral membrane proteins, and protein fragments.
Book Synopsis Plant ABC Transporters by : Markus Geisler
Download or read book Plant ABC Transporters written by Markus Geisler and published by Springer. This book was released on 2014-09-06 with total page 333 pages. Available in PDF, EPUB and Kindle. Book excerpt: This book is devoted to the fascinating superfamily of plant ATP-binding cassette (ABC) transporters and their variety of transported substrates. It highlights their exciting biological functions, covering aspects ranging from cellular detoxification, through development, to symbiosis and defense. Moreover, it also includes a number of chapters that center on ABC transporters from non-Arabidopsis species. ABC proteins are ubiquitous, membrane-intrinsic transporters that catalyze the primary (ATP-dependent) movement of their substrates through biological membranes. Initially identified as an essential aspect of a vacuolar detoxification process, genetic work in the last decade has revealed an unexpectedly diverse variety of ABC transporter substrates, which include not only xenobiotic conjugates, but also heavy metals, lipids, terpenoids, lignols, alkaloids and organic acids. The discovery that members of the ABCB and ABCG family are involved in the movement of phytohormones has further sparked their exploration and provided a new understanding of the whole family. Accordingly, the trafficking, regulation and structure-function of ABCB-type auxin transporters are especially emphasized in this book.
Book Synopsis Membrane Protein Assembly by : Gunnar von Heijne
Download or read book Membrane Protein Assembly written by Gunnar von Heijne and published by R. G. Landes. This book was released on 1997 with total page 300 pages. Available in PDF, EPUB and Kindle. Book excerpt:
Download or read book Pichia Protocols written by James M Cregg and published by Springer Science & Business Media. This book was released on 2007-08-08 with total page 553 pages. Available in PDF, EPUB and Kindle. Book excerpt: This book focuses on recent developments of Pichia pastoris as a recombinant protein production system. Highlighted topics include a discussion on the use of fermentors to grow Pichia pastoris, information on the O- and N-linked glycosylation, methods for labeling Pichia pastoris expressed proteins for structural studies, and the introduction of mutations in Pichia pastoris genes by the methods of restriction enzyme-mediated integration (REMI). Each chapter presents cutting-edge and cornerstone protocols for utilizing P. pastoris as a model recomibinant protein production system. This volume fully updates and expands upon the first edition.
Book Synopsis TRP Ion Channel Function in Sensory Transduction and Cellular Signaling Cascades by : Wolfgang B. Liedtke, MD, PH.D.
Download or read book TRP Ion Channel Function in Sensory Transduction and Cellular Signaling Cascades written by Wolfgang B. Liedtke, MD, PH.D. and published by CRC Press. This book was released on 2006-09-29 with total page 502 pages. Available in PDF, EPUB and Kindle. Book excerpt: Since the first TRP ion channel was discovered in Drosophila melanogaster in 1989, the progress made in this area of signaling research has yielded findings that offer the potential to dramatically impact human health and wellness. Involved in gateway activity for all five of our senses, TRP channels have been shown to respond to a wide range of st
Book Synopsis Biology of Serpins by : James Whisstock
Download or read book Biology of Serpins written by James Whisstock and published by Academic Press. This book was released on 2011-07-18 with total page 461 pages. Available in PDF, EPUB and Kindle. Book excerpt: Serpins are a group of proteins with similar structures that were first identified as a set of proteins able to inhibit proteases. The acronym serpin was originally coined because many serpins inhibit chymotrypsin-like serine proteases. This volume of Methods in Ezymology is split into 2 parts and comprehensively covers the subject.
Book Synopsis Insoluble Proteins by : Elena García-Fruitós
Download or read book Insoluble Proteins written by Elena García-Fruitós and published by Humana Press. This book was released on 2014-12-02 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: With insolubility proving to be one of the most crippling bottlenecks in the protein production and purification process, this volume serves to aid researchers working in the recombinant protein production field by describing a wide number of protocols and examples. Insoluble Proteins: Methods and Protocols includes chapters that describe not only the recombinant protein production in different expression systems but also different purification and characterization methods to finally obtain these difficult-to-obtain proteins. Beginning with protein production methods using both prokaryotic and eukaryotic expression systems, the book continues with purification protocols using insoluble proteins, the characterization of insoluble proteins, as well as a general overview of interesting applications of insoluble proteins. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Comprehensive and practical, Insoluble Proteins: Methods and Protocols aims to provide the scientific community with detailed and reliable state-of-the-art protocols that are used in order to successfully produce and purify recombinant proteins prone to aggregate.
Book Synopsis The Next Generation in Membrane Protein Structure Determination by : Isabel Moraes
Download or read book The Next Generation in Membrane Protein Structure Determination written by Isabel Moraes and published by Springer. This book was released on 2016-08-23 with total page 188 pages. Available in PDF, EPUB and Kindle. Book excerpt: This book reviews current techniques used in membrane protein structural biology, with a strong focus on practical issues. The study of membrane protein structures not only provides a basic understanding of life at the molecular level but also helps in the rational and targeted design of new drugs with reduced side effects. Today, about 60% of the commercially available drugs target membrane proteins and it is estimated that nearly 30% of proteins encoded in the human genome are membrane proteins. In recent years much effort has been put towards innovative developments to overcome the numerous obstacles associated with the structure determination of membrane proteins. This book reviews a variety of recent techniques that are essential to any modern researcher in the field of membrane protein structural biology. The topics that are discussed are not commonly found in textbooks. The scope of this book includes: Expression screening using fluorescent proteins The use of detergents in membrane protein research The use of NMR Synchrotron developments in membrane protein structural biology Visualisation and X-ray data collection of microcrystals X-ray diffraction data analysis from multiple crystals Serial millisecond crystallography Serial femtosecond crystallography Membrane protein structures in drug discovery The information provided in this book should be of interest to anyone working in the area of structural biology. Students will find carefully prepared overviews of basic ideas and advanced protein scientists will find the level of detail required to apply the material directly to their day to day work. Chapters 4, 5, 6, 8 and 9 of this book are published open access under a CC BY 4.0 license at link.springer.com.
Download or read book Lasso Peptides written by Yanyan Li and published by Springer. This book was released on 2014-10-21 with total page 113 pages. Available in PDF, EPUB and Kindle. Book excerpt: Lasso peptides form a growing family of fascinating ribosomally-synthesized and post-translationally modified peptides produced by bacteria. They contain 15 to 24 residues and share a unique interlocked topology that involves an N-terminal 7 to 9-residue macrolactam ring where the C-terminal tail is threaded and irreversibly trapped. The ring results from the condensation of the N-terminal amino group with a side-chain carboxylate of a glutamate at position 8 or 9, or an aspartate at position 7, 8 or 9. The trapping of the tail involves bulky amino acids located in the tail below and above the ring and/or disulfide bridges connecting the ring and the tail. Lasso peptides are subdivided into three subtypes depending on the absence (class II) or presence of one (class III) or two (class I) disulfide bridges. The lasso topology results in highly compact structures that give to lasso peptides an extraordinary stability towards both protease degradation and denaturing conditions. Lasso peptides are generally receptor antagonists, enzyme inhibitors and/or antibacterial or antiviral (anti-HIV) agents. The lasso scaffold and the associated biological activities shown by lasso peptides on different key targets make them promising molecules with high therapeutic potential. Their application in drug design has been exemplified by the development of an integrin antagonist based on a lasso peptide scaffold. The biosynthesis machinery of lasso peptides is therefore of high biotechnological interest, especially since such highly compact and stable structures have to date revealed inaccessible by peptide synthesis. Lasso peptides are produced from a linear precursor LasA, which undergoes a maturation process involving several steps, in particular cleavage of the leader peptide and cyclization. The post-translational modifications are ensured by a dedicated enzymatic machinery, which is composed of an ATP-dependent cysteine protease (LasB) and a lactam synthetase (LasC) that form an enzymatic complex called lasso synthetase. Microcin J25, produced by Escherichia coli AY25, is the archetype of lasso peptides and the most extensively studied. To date only around forty lasso peptides have been isolated, but genome mining approaches have revealed that they are widely distributed among Proteobacteria and Actinobacteria, particularly in Streptomyces, making available a rich resource of novel lasso peptides and enzyme machineries towards lasso topologies.
Book Synopsis Bacterial Persistence by : Jan Michiels
Download or read book Bacterial Persistence written by Jan Michiels and published by Humana. This book was released on 2015-10-15 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: This volume presents a comprehensive collection of methods that have been instrumental to the current understanding of bacterial persisters. Chapters in the book cover topics ranging from general methods for measuring persister levels in Escherichia coli cultures, protocols for the determination of the persister subpopulation in Candida albicans, quantitative measurements of Type I and Type II persisters using ScanLag, to in vitro and in vivo models for the study of the intracellular activity of antibiotics. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, Bacterial Persistence: Methods and Protocols brings together the most respected researchers in bacterial persistence whose studies will remain vital to understanding this field for many years to come.
Book Synopsis Expression, Purification, and Structural Biology of Membrane Proteins by : Camilo Perez
Download or read book Expression, Purification, and Structural Biology of Membrane Proteins written by Camilo Perez and published by Humana. This book was released on 2021-03-14 with total page 423 pages. Available in PDF, EPUB and Kindle. Book excerpt: This book collects up-to-date advanced protocols and advice from leading experts in the area of membrane protein biology that can be applied to structural and functional studies of any membrane protein system. The contents explore methods for cloning and expression of membrane proteins and membrane protein complexes in prokaryotic and eukaryotic systems, approaches for protein purification, nanobody applications, as well as biophysical characterization and much more. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and thorough, Expression, Purification, and Structure Biology of Membrane Proteins serves to guide and encourage young researchers and newcomers to the field to tackle bold new studies on membrane proteins. Chapter 11 is available open access under a CC-BY 4.0 license via link.springer.com.