Protein Self-Assembly

This volume explores experimental and computational approaches to measuring the most widely studied protein assemblies, including condensed liquid phases, aggregates, and crystals. The chapters in this book are organized into three parts: Part One looks at the techniques used to measure protein-protein interactions and equilibrium protein phases in dilute and concentrated protein solutions; Part Two describes methods to measure kinetics of aggregation and to characterize the assembled state; and Part Three details several different computational approaches that are currently used to help researchers understand protein self-assembly. 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.

Thorough and cutting-edge, Protein Self-Assembly: Methods and Protocols is a valuable resource for researchers who are interested in learning more about this developing field.

Includes cutting-edge methods and protocols Provides step-by-step detail essential for reproducible results Contains key notes and implementation advice from the experts

Contenu

Measuring Non-Specific Protein-Protein Interactions by Dynamic Light Scattering.- Light Scattering to Quantify Protein-Protein Interactions at High Protein Concentrations.- Quantitative Evaluation of Protein Solubility in Aqueous Solutions by PEG-Induced Liquid-Liquid Phase Separation.- Measuring Protein Solubility.- Integral caa3-Cytochrome c Oxidase from Thermus thermophiles: Purification and Crystallization.- Aggregation Profiling of C9orf72 Dipeptide Repeat Proteins Transgenically Expressed in Drosophila melanogaster using an Analytical Ultracentrifuge Equipped with Fluorescence Detection.- Sample Preparation and Size Analysis of C9orf72 Dipeptide Repeat Proteins Expressed in Drosophila melanogaster using Semi-Denaturing Agarose Gel Electrophoresis.- The Use of High Performance Liquid Chromatography for the Characterization of the Unfolding and Aggregation of Dairy Proteins.- Differential Scanning Calorimetry to Quantify Heat Induced Aggregation in Concentrated Protein Solution.- Nanoparticle Tracking Analysis to Examine the Temperature-Induced Aggregation of Proteins.- Evaluation of Temporal Aggregation Processes using Spatial Intensity Distribution Analysis.- Fluorescence Correlation Spectroscopy for Particle Sizing in Highly Concentrated Protein Solutions.- Size Determination of Protein Oligomers/Aggregates Using Diffusion NMR Spectroscopy.- Patchy Particle models to Understand Protein Phase Behavior.- Obtaining Soft Matter Models of Proteins and their Phase Behavior.- Binding Free Energies of Conformationally Disordered Peptides through Extensive Sampling and End-Point Methods.- Atomistic Simulation Tools to Study Protein Self-Aggregation.