Membrane Protein Protocols

Author: Barry Steven Selinsky
Publisher: Springer Science & Business Media
ISBN: 159259400X
Size: 78.99 MB
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A collection of key techniques for the study of receptors and transport proteins. The book provides examples of how different membrane proteins can be over-expressed in both prokaryotic and eukaryotic expression systems, how natural and overexpressed proteins can be solubilized from their host membranes, and how the solubilized protein can be purified in active form. Each protocols contains step-by-step instructions to ensure success, troubleshooting advice, lists of reagents, and tips on avoiding pitfalls.

Membrane Protein Purification And Crystallization

Author: Carola Hunte
Publisher: Elsevier
ISBN: 0080536174
Size: 66.82 MB
Format: PDF, Docs
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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

Membrane Protein Structure Determination

Author: Jean-Jacques Lacapère
Publisher: Humana PressInc
ISBN:
Size: 57.38 MB
Format: PDF
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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. --

Membrane Protein Protocols

Author: Barry S. Selinsky
Publisher: Humana Press
ISBN: 9781588291240
Size: 33.17 MB
Format: PDF, Docs
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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.

Membrane Protein Structure And Function Characterization

Author: Jean-Jacques Lacapere
Publisher: Humana Press
ISBN: 9781493971497
Size: 61.98 MB
Format: PDF
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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.

Heterologous Expression Of Membrane Proteins

Author: Isabelle Mus-Veteau
Publisher: Humana Press
ISBN: 9781493936359
Size: 27.30 MB
Format: PDF, Kindle
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This detailed volume encompasses chapters from leading experts in the area of membrane proteins who describe step-by-step protocols developed these last few years to improve the functional production and stabilization of recombinant integral membrane proteins (IMPs). Membrane proteins play a key role in numerous pathologies such as cancer, cystic fibrosis, epilepsy, hyperinsulinism, and Alzheimer’s disease, yet studies on these and other disorders are hampered by a lack of information about the proteins involved. This book sets out to aid researchers in rectifying this situation. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, 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, Second Edition serves as an ideal guide for scientists attempting to delve deeper into the myriad unique IMP structures.

The Protein Protocols Handbook

Author: John M. Walker
Publisher: Springer Science & Business Media
ISBN: 1603272593
Size: 14.14 MB
Format: PDF, Docs
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In The Protein Protocols Handbook, I have attempted to provide a cross-section of analytical techniques commonly used for proteins and peptides, thus providing a benehtop manual and guide both for those who are new to the protein chemistry laboratory and for those more established workers who wish to use a technique for the first time. We each, of course, have our own favorite, commonly used gel system, g- staining method, blotting method, and so on; I'm sure you will find yours here. H- ever, I have also described a variety of altematives for many of these techniques; though they may not be superior to the methods you commonly use, they may nev- theless be more appropriate in a particular situation. Only by knowing the range of techniques that are available to you, and the strengths and limitations of these te- niques, will you be able to choose the method that best suits your purpose.

Protein Purification Protocols

Author: Paul Cutler
Publisher: Springer Science & Business Media
ISBN: 159259655X
Size: 29.29 MB
Format: PDF
View: 2020
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This new edition of Protein Purification Protocols completely updates the existing protocols to reflect recent advances and adds the enormous new array of proteomic techniques for protein isolation and analysis. These cutting-edge techniques include not only two-dimensional gel electrophoresis for analysis and characterization, but also analytical chromatography for multidimensional separations of proteins and peptides, and mass spectrometry for isolating proteins.

Protein Purification Protocols

Author: Shawn Doonan
Publisher: Springer Science & Business Media
ISBN: 1592595456
Size: 71.35 MB
Format: PDF, ePub, Docs
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A comprehensive collection of essential, time-tested recipes for successful protein fractionation and purification in any experimental circumstance. The protocols give step-by-step instructions on how to select a source for the protein of interest, how to obtain a usable initial extract, how to purify the protein from that extract using both chemical and molecular methods, and how to dry and store the purified protein. Protein Purification Protocols provides all that is needed to design and carry out a successful purification program. It helps both experienced and novice investigators to clarify and define their purification problems and then provides a comprehensive set of tools for a practical solution.

Electron Crystallography Of Soluble And Membrane Proteins

Author: Ingeborg Schmidt-Krey
Publisher: Humana Press
ISBN: 9781493958764
Size: 78.59 MB
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The basic principle of electron crystallography is to calculate a 3D density map by combining the amplitudes obtained from electron diffraction patterns with the experimental phases calculated from images of two-dimensional crystals of membrane or soluble proteins. This technology is very well developed and has produced a number of atomic models of membrane proteins in a lipid environment. Focused on comprehensive experimental protocols, Electron Crystallography of Soluble and Membrane Proteins: Methods and Protocols covers the entire range of techniques used in electron crystallography, including protein sample preparation, 2D crystallization, and screening in negative stain over electron cryo-microscopy (cryo-EM) and data processing, as well as modeling of conformational changes. Additional chapters provide perspective on past, present, and future challenges as well as complementary methods. Written for the popular Methods in Molecular BiologyTM series, the work contains the kind of detailed descriptions and implementation advice necessary to ensure successful results. Comprehensive and cutting-edge, Electron Crystallography of Soluble and Membrane Proteins: Methods and Protocols serves laboratories new to the methods as well as state-of-the-art facilities pursuing this exciting area of protein science.