Membrane Protein Protocols

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Membrane Protein Protocols

Author : Barry S. Selinsky
Publisher : Springer Science & Business Media
Page : 330 pages
File Size : 41,5 Mb
Release : 2008-02-03
Category : Science
ISBN : 9781592594009

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Membrane Protein Protocols by Barry S. Selinsky Pdf

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.

Heterologous Expression of Membrane Proteins

Author : Isabelle Mus-Veteau
Publisher : Springer Nature
Page : 472 pages
File Size : 52,5 Mb
Release : 2022-06-30
Category : Science
ISBN : 9781071623688

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Heterologous Expression of Membrane Proteins by Isabelle Mus-Veteau Pdf

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.

Membrane Protein Structure and Function Characterization

Author : Jean-Jacques Lacapere
Publisher : Humana
Page : 419 pages
File Size : 51,7 Mb
Release : 2018-06-10
Category : Science
ISBN : 1493983997

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Membrane Protein Structure and Function Characterization by Jean-Jacques Lacapere Pdf

In this present volume, different approaches are detailed to produce membrane proteins, purify them, study their function, determine their structure, and model them in membrane. Since every membrane protein behaves mostly in a unique way /fashion, knowledge of guidelines and tricks may help to increase chances to express, purify and characterize a peculiar membrane protein. Production of correctly folded protein remains a challenge. Moreover, getting a functional and stable protein requires to optimize membrane mimicking environments that can be detergent or artificial membranes. In some cases, the finding of the correct ligand which will stabilize the desired conformation is needed. In other cases, stabilization can be obtained using specific antibodies. This volume also presents different techniques to analyze the functional status of membrane proteins. Written in the highly successful Methods in Molecular Biology series format, chapters in Membrane Protein Structure and Function Characterization: Methods and Protocols provide different techniques to analyze the functional and structural status of membrane proteins. 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, Membrane Protein Structure and Function Characterization: Methods and Protocols aims to ensure successful results in the further study of this vital field.

Membrane Protein Purification and Crystallization

Author : Carola Hunte,Gebhard von Jagow,Hermann Schagger
Publisher : Elsevier
Page : 339 pages
File Size : 55,8 Mb
Release : 2003-01-05
Category : Science
ISBN : 9780080536170

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Membrane Protein Purification and Crystallization by Carola Hunte,Gebhard von Jagow,Hermann Schagger Pdf

This second edition of Membrane Protein Purification and Crystallization, A Practical Guide is written for bench scientists working in the fields of biochemistry, biology, and proteomic research. This guide presents isolation and crystallization techniques in a concise form, emphasizing the critical aspects unique to membrane proteins. It explains the principles of the methods and provides protocols of general use, permitting researchers and students new to this area to adapt these techniques to their particular needs. This edition is not only an update but is comprised mainly of new contributions. It is the first monograph compiling the essential approaches for membrane protein crystallization, and emphasizes recent progress in production and purification of recombinant membrane proteins. Provides general guidelines and strategies for isolation and crystallization of membrane proteins Gives detailed protocols that have wide application, and low specialized equipment needs Emphasizes recent progress in production and purification of recombinant membrane proteins, especially of histidine-tagged and other affinity-epitope-tagged proteins Summarizes recent developments of Blue-Native PAGE, a high resolution separation technique, which is independent of the use of recombinant techniques, and is especially suited for proteomic analyses of membrane protein complexes Gives detailed protocols for membrane protein crystallization, and describes the production and use of antibody fragments for high resolution crystallization Presents a comprehensive guide to 2D-crystallization of membrane proteins

Biophysics of Membrane Proteins

Author : Vincent L. G. Postis,Adrian Goldman
Publisher : Humana
Page : 336 pages
File Size : 52,8 Mb
Release : 2022-02-28
Category : Science
ISBN : 107160726X

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Biophysics of Membrane Proteins by Vincent L. G. Postis,Adrian Goldman Pdf

This volume provides recent advances in the field of biophysics of membrane proteins. Chapters are divided into several parts: detailing biochemistry and functional analysis, experimental and theoretical structural determinations, membrane protein dynamics, and conformation studies. 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, Biophysics of Membrane Proteins: Methods and Protocols aims to provide comprehensive protocols with notes to help further the understanding of key membrane protein structure and function for students, academics, and industrial researchers.

A Practical Guide to Membrane Protein Purification

Author : Gebhard von Jagow
Publisher : Elsevier
Page : 185 pages
File Size : 52,8 Mb
Release : 2013-10-22
Category : Science
ISBN : 9780080571720

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A Practical Guide to Membrane Protein Purification by Gebhard von Jagow Pdf

A Practical Guide to Membrane Protein Purification is written especially for researchers who have some familarity with separation of water-soluble proteins, but who may not be aware of the pitfalls they face with membrane proteins. This guide presents techniques in a concise form, emphasizing the aspects unique to membrane proteins. The book explains the principles of the methods, permitting researchers and students new to this area to adapt these techniques to their particular needs. The second volume in the series, this book is an essential manual for investigations of structure and function of native membrane proteins, as well as for purification of these proteins for immunization and protein sequencing. Separation, Detection, and Characterization of Biological Macromolecules is a new series of laboratory guides. Each volume focuses on a topic of central interest to scientists and students in biomedical and biological research. Introductory chapters are followed by clear, step-by-step protocols that present principles and practice. These concise manuals are designed for optimal understanding of methods as well as for practical benchtop use. Provides general guidelines and strategies for isolation of membrane proteins Describes detailed practical procedures that have been the widest applications, and lowest specialized equipment needs Gives special emphasis to new native and denaturing electrophoresis techniques Explains modifications of techniques used for water-soluble proteins

Protein Purification Protocols

Author : Shawn Doonan
Publisher : Springer Science & Business Media
Page : 406 pages
File Size : 45,7 Mb
Release : 2008-02-02
Category : Science
ISBN : 9781592595457

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Protein Purification Protocols by Shawn Doonan Pdf

Hans Neurath has written that this is the second golden era of enzymology {Protein Science [1994], vol. 3, pp. 1734—1739); he could with justice have been more general and referred to the second golden age of protein chemistry. The last two decades have seen enormous advances in our understanding of the structures and functions of pro teins arising on the one hand from improvements and developments in analytical techniques {see the companion volume, Basic Protein and Peptide Protocols, in this series) and on the other hand from the tech nologies of molecular genetics. Far from turning the focus away from protein science, the ability to isolate, analyze, and express genes has increased interest in proteins as gene products. Hence, many laborato ries are now getting involved in protein isolation for the first time, either as an essential adjunct to their work in molecular genetics or because of a curiosity to know more about the products of the genes that they have been studying. Protein Purification Protocols is aimed mainly at these newcom ers to protein purification, but it is hoped that it will also be of value to established practitioners who may find here techniques that they have not tried, but which might well be most applicable in their work. With the exception mainly of the first and last chapters, the format of the contributions to the present book conform to the established format of the Methods in Molecular Biology series.

Expression, Purification, and Structural Biology of Membrane Proteins

Author : Camilo Perez,Timm Maier
Publisher : Humana
Page : 423 pages
File Size : 43,9 Mb
Release : 2021-03-14
Category : Science
ISBN : 1071603752

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Expression, Purification, and Structural Biology of Membrane Proteins by Camilo Perez,Timm Maier Pdf

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.

Membrane Protein Structure Determination

Author : Jean-Jacques Lacapère
Publisher : Methods in Molecular Biology
Page : 482 pages
File Size : 54,7 Mb
Release : 2010-08-06
Category : Science
ISBN : STANFORD:36105211750646

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Membrane Protein Structure Determination by Jean-Jacques Lacapère Pdf

Membrane proteins, representing nearly 40% of all proteins, are key components of cells involved in many cellular processes, yet only a small number of their structures have been determined. Membrane Protein Structure Determination: Methods and Protocols presents many detailed techniques for membrane protein structure determination used today by bringing together contributions from top experts in the field. Divided into five convenient sections, the book covers various strategies to purify membrane proteins, approaches to get three dimensional crystals and solve the structure by x-ray diffraction, possibilities to gain structural information for a membrane protein using electron microscopy observations, recent advances in nuclear magnetic resonance (NMR), and molecular modelling strategies that can be used either to get membrane protein structures or to move from atomic structure to a dynamic understanding of a molecular functioning mechanism. Written in the highly successful Methods in Molecular BiologyTM 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 easy to use, Membrane Protein Structure Determination: Methods and Protocols serves as an ideal reference for scientists seeking to further our knowledge of these vital and versatile proteins as well as our overall understanding of the complicated world of cell biology.

Biophysics of Membrane Proteins

Author : Vincent L. G. Postis,Adrian Goldman
Publisher : Humana
Page : 0 pages
File Size : 51,7 Mb
Release : 2021-02-14
Category : Science
ISBN : 1071607235

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Biophysics of Membrane Proteins by Vincent L. G. Postis,Adrian Goldman Pdf

This volume provides recent advances in the field of biophysics of membrane proteins. Chapters are divided into several parts: detailing biochemistry and functional analysis, experimental and theoretical structural determinations, membrane protein dynamics, and conformation studies. 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, Biophysics of Membrane Proteins: Methods and Protocols aims to provide comprehensive protocols with notes to help further the understanding of key membrane protein structure and function for students, academics, and industrial researchers.

Membrane Protein Structure and Dynamics

Author : Nagarajan Vaidehi,Judith Klein-Seetharaman
Publisher : Humana Press
Page : 357 pages
File Size : 44,8 Mb
Release : 2012-09-14
Category : Science
ISBN : 1627030247

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Membrane Protein Structure and Dynamics by Nagarajan Vaidehi,Judith Klein-Seetharaman Pdf

Membrane proteins play key roles in numerous cellular processes, in particular mediating cell-to-cell communication and signaling events that lead to a multitude of biological effects. Membrane proteins have also been implicated in many critical diseases such as atherosclerosis, hypertension, diabetes and cancer. In Membrane Protein Structure Predictions Methods: Methods and Protocols, expert researcher in the field detail the advances in both experimental and computational approaches of the structure, dynamics and interactions of membrane proteins dividing the volume into two sections. The first section details the procedures used for measurements of structure and dynamics of membrane proteins. While the second section contains a survey of the computational methods that have played a critical role in membrane protein structure prediction as well as in providing atomic level insight into the mechanism of the dynamics of membrane receptors. Written in the highly successful Methods in Molecular BiologyTM series format, the chapters include the kind of detailed description and implementation advice that is crucial for getting optimal results in the laboratory. Thorough and intuitive, Membrane Protein Structure Predicitons: Methods and Protocols seeks to aid scientists in the further study of membrane protein structure and function.

Production of Membrane Proteins

Author : Anne Skaja Robinson
Publisher : John Wiley & Sons
Page : 631 pages
File Size : 53,8 Mb
Release : 2011-06-15
Category : Science
ISBN : 9783527634538

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Production of Membrane Proteins by Anne Skaja Robinson Pdf

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.

Membrane Biogenesis

Author : Doron Rapaport,Johannes M. Herrmann
Publisher : Humana
Page : 0 pages
File Size : 53,7 Mb
Release : 2016-08-23
Category : Science
ISBN : 149396304X

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Membrane Biogenesis by Doron Rapaport,Johannes M. Herrmann Pdf

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.

Membrane Proteomics

Author : Matthew J. Peirce,Robin Wait
Publisher : Humana Press
Page : 0 pages
File Size : 45,6 Mb
Release : 2010-11-19
Category : Science
ISBN : 1617378895

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Membrane Proteomics by Matthew J. Peirce,Robin Wait Pdf

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.

Structure and Function of Membrane Proteins

Author : Ingeborg Schmidt-Krey,James C. Gumbart
Publisher : Humana
Page : 358 pages
File Size : 50,7 Mb
Release : 2021-04-21
Category : Science
ISBN : 1071613936

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Structure and Function of Membrane Proteins by Ingeborg Schmidt-Krey,James C. Gumbart Pdf

This book examines detailed experimental and computational approaches for the analysis of many aspects vital to the understanding of membrane protein structure and function. Readers will receive guidance on the selection and use of methods for over-expression and purification, tools to characterize membrane proteins within different phospholipid bilayers, direction on functional studies, and approaches to determine the structures of membrane proteins. Detailed experimental steps for specific membrane proteins with critical notes allow the protocols to be modified to different systems. Written for the highly successful Methods in Molecular Biology series, chapters include the kind of practical information and implementation advice that leads to excellent, reproducible results. Authoritative and up-to-date, Structure and Function Studies of Membrane Proteins serves as an ideal guide for biologists, biochemists, and biophysicists striving to further understand these essential proteins and their many biological roles.