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There have been many important advances in our understanding of biological membrane structure and function over the last decade. Much of this progress has been driven by the development of new techniques for studying membrane components and their interactions. Tradition ally, the investigation of membranes has largely occurred within the domains of biochemistry, physical chemistry, and cell biology; but many of the most significant advances have resulted from an expansion into other disciplines, such as molecular biology, immunology, and the clinical sciences. In these two volumes we have attempted to bring together some of these techniques—to combine the old and the new. Biomembrane Protocols: II. Architecture and Function is concerned exclusively with the architecture and activities of membranes, whereas the companion volume, Biomembrane Protocols: I. Isolation and Analysis, deals with their isolation and compositional analysis. The aim of each chapter is to provide detailed technical and meth odological information that will allow the reader to perform the tech nique successfully, without the need to consult other texts. Detailed reviews of membrane structure and function are not included, except where they are relevant to the choice or efficacy of a particular proce dure. In addition to Materials and Methods sections, each chapter has a Notes section that explains the reasons for taking certain steps and pro vides practical tips for applying the techniques to other systems.
This second volume of protocols offers the most comprehensive set of modern analytical techniques available for studying the architecture and function of membranes. It features the application of biochemical, spectroscopic, and fluorimetric methods to the investigation of molecular topology, the dynamic aspects of membrane structure, and ion transport. Antibody technology, analysis of molecules involved in intracellular signaling, and receptor-ligand interactions are also covered.
Expert practitioners combine the traditional basic techniques of membrane isolation and analysis with modern advances in molecular biology, immunology, and clinical sciences to devise analytical techniques that are successful for expert and novice alike. The techniques are used in the isolation and compositional analysis of membranes and address animal, plant, and microbial cells. The topics covered in this first volume include the isolation of subcellular fractions, centrifugation, the extraction and quantitation of lipids, the separation and analysis of proteins, and the isolation and characterization of glycosylated molecules, glycoproteins, proteoglycans, and glycolipids.
The first volume of a comprehensive two-volume collection of methodologies for studying membrane components and their interactions, gleaned from the areas of biochemistry, physical chemistry, cell biology, molecular biology, immunology, and clinical sciences. Volume 1 is concerned exclusively with the isolation and compositional analysis of membranes, while Volume 2 will deal with their architecture and activities. Experts in the field provide step-by-step technical and methodological information to enable successful performance of each technique. Plastic comb binding. Annotation copyright by Book News, Inc., Portland, OR
Membrane proteins and membrane lipids form complex interactive systems that are highly dynamic and able to be studied only by combinations of different in vivo and in vitro techniques. In Membrane Biogenesis: Methods and Protocols, experts in the field present a broad collection of methods to study the biogenesis and function of cellular membranes. Beginning with how membrane lipids or membrane proteins can be studied, this detailed volume continues with sections covering different procedures to investigate the interaction of membrane proteins among each other or with membrane lipids, methods to study the biogenesis of membrane proteins and the dynamics of organelles, as well as protocols for the analyses of the functions or complex organization of membrane proteins. Written in the highly successful Methods in Molecular BiologyTM series format, chapters include introductions to their respective topics, lists of materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Extensive and easily applicable, Membrane Biogenesis: Methods and Protocols provides readers with a comprehensive but still concise collection including both basic protocols of rather general application and more specialized methods for specific and novel techniques.
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.
There have been many important advances in our understanding of biological membrane structure and function over the last decade. Much of this progress has been driven by the development of new techniques for studying membrane components and their interactions. Tradition ally, the investigation of membranes has largely occurred within the domains of biochemistry, physical chemistry, and cell biology; but many of the most significant advances have resulted from an expansion into other disciplines, such as molecular biology, immunology, and the clinical sciences. In these two volumes we have attempted to bring together some of these techniques—to combine the old and the new. Biomembrane Protocols: II. Architecture and Function is concerned exclusively with the architecture and activities of membranes, whereas the companion volume, Biomembrane Protocols: I. Isolation and Analysis, deals with their isolation and compositional analysis. The aim of each chapter is to provide detailed technical and meth odological information that will allow the reader to perform the tech nique successfully, without the need to consult other texts. Detailed reviews of membrane structure and function are not included, except where they are relevant to the choice or efficacy of a particular proce dure. In addition to Materials and Methods sections, each chapter has a Notes section that explains the reasons for taking certain steps and pro vides practical tips for applying the techniques to other systems.
This detailed book explores examples of current in vitro and in silico techniques that are at the forefront of lipid membrane research today. Beginning with methods and strategies associated with the creation and use of lipid membrane models in various research settings, the volume continues with electrical impedance spectroscopy strategies and methods to identify how ions and proteins interact with model lipid bilayers, guidance on lipid bilayer in silico molecular dynamics modeling, novel techniques to explore lipid bilayer characteristics using neutron scattering, IR spectroscopy, and atomic force microscopy (AFM), as well as unique fluorescence techniques. Written in the highly successful Methods in Molecular Biology series style, chapters include introductions to their respective topics, lists of the necessary materials, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, Membrane Lipids: Methods and Protocols serves as an ideal guide for researchers seeking to further investigate the often complicated world of lipid membrane biophysics.
The membranes surrounding cells and organelles constitute their interface with the local environment. The functions of membrane proteins include cell/cell and cell/extracellular matrix recognition, the reception and transduction of extracellular signals, and the tra- port of proteins, solutes and water molecules. Abnormal membrane protein expression has profound biological effects and may, for example, underlie phenotypic and functional differences between normal and tumour cells. Moreover the accessibility, particularly of plasma proteins traversing the plasma membrane of cells, makes them of particular ut- ity to the therapeutic intervention in disease. Indeed, it is estimated that of all currently licensed pharmaceuticals, approximately 70% target proteins resident in the plasma m- brane. In theory, unbiased technologies such as proteomics have the power to de?ne patterns of membrane protein expression characteristic of distinct states of cellular development, differentiation or disease, and thereby identify novel markers of, or targets for intervention in, disease. However, although about 25% of open reading frames in fully sequenced genomes are estimated to encode integral membrane proteins, global analysis of membrane protein expression has proved problematic. Membrane protein analysis poses unique challenges at the level of extraction, solubilization, and separation in particular, and to a lesser extent of identi?cation and quantitation. These challenges have, however, fostered creativity, in- vation, and technical advances, many of which are brought together in Membrane P- teomics.