Multiprotein Complexes : Methods and Protocols / edited by Arnaud Poterszman.

Format:

Book

Contributor:

Poterszman, Arnaud, Editor.

SpringerLink (Online service)

Series:

Springer Protocols (Springer-12345)

Methods in molecular biology 1940-6029 ; 2247

Methods in Molecular Biology, 1940-6029 ; 2247

Language:

English

Subjects (All):

Proteins.

Protein Biochemistry.

Local Subjects:

Protein Biochemistry.

Physical Description:

1 online resource (XII, 340 pages) : 82 illustrations, 70 illustrations in color.

Edition:

1st ed. 2021.

Contained In:

Springer Nature eBook

Place of Publication:

New York, NY : Springer US : Imprint: Humana, 2021.

System Details:

text file PDF

Summary:

This volume explores strategies and detailed protocols for the preparation of macromolecular complexes and their characterization in view of structural analysis. The chapters in this book are separated into three parts: Part One focuses on sample preparation, and covers strategies for recombinant expression of multiprotein complexes in prokaryotic and eukaryotic hosts, for genome engineering using the CRISPR/Cas9 system and for production of specific binders such as reformatted antibodies and artificial binding proteins. Part Two looks at the biophysical methods that can provide useful indicators for sample optimization, and often complement structural information obtained with core technologies for structure determination-x-ray crystallography and cryo-electron microscopy-by quantitative solution data. Part Three discusses the characterization of multiprotein complexes in a cellular environment using the latest technologies and in vivo approaches. 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. Cutting-edge and authoritative, Multiprotein Complexes: Methods and Protocols is a valuable resource for structural and molecular biologists who need to prepare multi-components for their applications, and for other scientists working on macromolecular assemblies from other angles that need to know the latest approaches that the field has to offer.

Contents:

Production of Multi-Subunit Membrane Protein Complexes

Production of Multiprotein Complexes using the Baculovirus Expression System: Homology Based and Restriction Free Cloning Strategies for Construct Design

Tagging Proteins with Fluorescent Reporters using the CRISPR/Cas9 System and Double Stranded DNA Donors

Validation of the Production of Antibodies in Different Formats in the HEK 293 Transient Gene Expression System

The Intervening Removable Affinity Tag (iRAT) System for the Production of Recombinant Antibody Fragments

Isolation of Artificial Binding Proteins (Affimer Reagents) for Use in Molecular and Cellular Biology

Interactions Measurements of Protein-DNA Complexes by Isothermal Titration Calorimetry (ITC) and Microscale Thermophoresis (MST)

switchSENSE Technology for Analysis of DNA Polymerase Kinetics

Sedimentation Velocity Methods for Characterization of Protein Heterogeneity and Protein Affinity Interactions

Hands on Native Mass Spectrometry Analysis of Multi-Protein Complexes

Studying Protein-DNA Interactions by Hydrogen/Deuterium Exchange Mass Spectrometry

Integrative Mass Spectrometry-Based Approaches for Modeling Macromolecular Assemblies

Optimization of Sample Preparation for the Observation of Macromolecular Complexes by Electron (cryo-)Microscopy

Solubilization and Stabilization of Native Membrane Proteins for Drug Discovery

Practical Aspects of Super-Resolution Imaging and Segmentation of Macromolecular Complexes by dSTORM

Multicolor FRET-FLIM Microscopy to Analyze Multiprotein Interactions in Live Cells

Context-Specific and Proximity-Dependent Labeling for the Proteomic Analysis of Spatiotemporally Defined Protein Complexes with Split-BioID

A Directed Evolution System for Lysine Deacetylases.

Other Format:

Printed edition:

ISBN:

978-1-0716-1126-5

9781071611265

Access Restriction:

Restricted for use by site license.