Intrinsically Disordered Proteins - Methods and Protocols

The edition details methods to study intrinsically disordered proteins (IDPs) including recent topics such as extremely high-affinity disordered complexes, kinetics that evade established concepts, liquid-liquid phase separation, and novel disorder-driven allosteric mechanisms. 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, Intrinsically Disordered Proteins: Methods and Protocols aims to help scientists with different backgrounds to further their investigations into these fascinating and dynamic molecules.
Chapter 24 is available open access under a CC BY 4.0 license via link.springer.com.
Chapters "40 and 42 " are available open access under a Creative Commons Attribution 4.0 International License via link.springer.com.

List of contents

Disorder for Dummies: Functional Mutagenesis of Transient Helical Segments in Disordered Proteins.- Computational Prediction of Intrinsic Disorder in Protein Sequences with the disCoP Meta-predictor.- Computational Prediction of Disordered Protein Motifs using SLiMSuite.- How to Annotate and Submit a Short Linear Motif to the Eukaryotic Linear Motif Resource.- Analyzing the Sequences of Intrinsically Disordered Regions with CIDER and localCIDER.- Exploring Protein Intrinsic Disorder with MobiDB.- An Easy Protocol for Evolutionary Analysis of Intrinsically Disordered Proteins.- Expression and Purification of an Intrinsically Disordered Protein.- Production of Intrinsically Disordered Proteins for Biophysical Studies; Tips and Tricks.- Recombinant Production of Monomeric Isotope-Enriched Aggregation-Prone Peptides: Polyglutamine Tracts and Beyond.- Cell-free Protein Synthesis of Small Intrinsically Disordered Proteins for NMR Spectroscopy.- Structural Analyses of Intrinsically Disordered Proteins by Small-Angle X-ray Scattering.- Determining Rg of IDPs from SAXS data.- Obtaining Hydrodynamic Radii of Intrinsically Disordered Protein Ensembles by Pulsed Field Gradient NMR Measurements.- Quantitative Protein Disorder Assessment using NMR Chemical Shifts.- Determination of pK_a Values in Intrinsically Disordered Proteins.- Paris-DÉCOR: A Protocol for the Determination of Fast Protein Backbone Amide Hydrogen Exchange Rates.- Predicting Conformational Properties of Intrinsically Disordered Proteins from Sequence.- Enhanced Molecular Dynamics Simulations of Intrinsically Disordered Proteins.- Computational Protocol for Determining Conformational Ensembles of Intrinsically Disordered Proteins.- Computing, Analyzing and Comparing the Radius of Gyration and Hydrodynamic Radius in Conformational Ensembles of Intrinsically Disordered Proteins.- Binding Thermodynamics to Intrinsically Disordered Protein Domains.- Analysis of Multivalent IDP Interactions: Stoichiometry, Affinity, and Local Concentration Effect Measurements.- NMR Lineshape Analysis of Intrinsically Disordered Protein Interactions.- Measuring Effective Concentrations Enforced by Intrinsically Disordered Linkers.- Determining the Protective Activity of IDPs under Partial Dehydration and Freeze-thaw Conditions.- Screening Intrinsically Disordered Regions for Short Linear Binding Motifs.- Probing IDP Interactions with Membranes by Fluorescence Spectroscopy.- Protocol for Investigating the Interactions between Intrinsically Disordered Proteins and Membranes by Neutron Reflectometry.- Interactions of IDPs with Membranes Using Dark State Exchange NMR Spectroscopy.- Determination of Binding Kinetics of Intrinsically Disordered Proteins by Surface Plasmon Resonance.- Measuring and Analysing Binding Kinetics of Coupled Folding and BindingReactions under Pseudo-first Order Conditions.- Understanding Binding Induced Folding by Temperature Jump.- Determining Binding Kinetics of Intrinsically Disordered Proteins by NMR spectroscopy.- Determination of Protein Phase Diagrams by Centrifugation.- In vitro Transition Temperature Measurement of Phase Separating Proteins by Microscopy.- Walking along a Protein Phase Diagram to Determine Coexistence Points by Static Light Scattering.- Expression and Purification of Intrinsically Disordered Abeta Peptide and Setup of Reproducible Aggregation Kinetics Experiment.- Measuring Interactions between Tau and Aggregation Inducers with Single Molecule Förster Resonance Energy Transfer.- Detection of Multisite Phosphorylation of Intrinsically Disordered Proteins using Phos-tag SDS-PAGE.- Multiple Site-specific Phosphorylation of IDPs Monitored by NMR.- Detection of Multisite Phosphorylation of Intrinsically Disordered Proteins usingQuantitative Mass-Spectrometry.- Targeting an Intrinsically Disordered Protein by Covalent Modification.- Recording in-cell NMR-spectra in Living Mammalian Cells.- In-cell NMR of Intrinsically Disordered Proteins in Mammalian Cells.- Analyzing IDPs in Interactomes. 

Product details

• Assisted by: Birth B Kragelund (Editor), Birthe B Kragelund (Editor), Birthe B. Kragelund (Editor), Skriver (Editor), Skriver (Editor), Karen Skriver (Editor)
• Publisher: Springer, Berlin
• Languages: English
• Product format: Paperback / Softback
• Released: 05.08.2021
• EAN: 9781071605264
• ISBN: 978-1-0-7160526-4
• No. of pages: 951
• Dimensions: 178 mm x 52 mm x 254 mm
• Illustrations: XVIII, 951 p. 231 illus., 192 illus. in color.
• Series: Methods in Molecular Biology
• Subject: Natural sciences, medicine, IT, technology > Biology > Biochemistry, biophysics