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.
Inhaltsverzeichnis
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 Aß 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.
