Protein Complex Assembly

This volume details the importance of multiple experimental techniques and computational methods needed to obtain the comprehensive picture of protein complex structure, dynamics and assembly afforded by the emerging field of integrative structural biology. Chapters guide readers through the broad spectrum of approaches required for a complete representation of protein complexes, including expression and purification, experimental characterization of structure and assembly, and computational methods for identifying protein complexes and modelling their 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.

Authoritative and cutting-edge, Protein Complex Assembly: Methods and Protocols aims to ensure successful results in the further study of this vital 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

Inhalt
Experimental Characterisation Of Protein Complex Structure, Dynamics, and Assembly.- High-Throughput Electron Cryo-Tomography of Protein Complexes and their Assembly.- Preparation of Tunable Microchips to Visualize Native Protein Complexes for Single Particle Electron Microscopy.- Time-resolved Cryo-Electron Microscopy Using a Microfluidic Chip.- Characterizing Protein-protein Interactions using Solution NMR Spectroscopy.- Reconstitution of Isotopically Labeled Ribosomal Protein L29 in the 50S Large Ribosomal Subunit For Solution-state and Solid-state NMR.- Characterizing Protein-Protein Interactions using Deep Sequencing Coupled to Yeast Surface Display.- Structurally Guided In Vivo Crosslinking.- Characterising Intact Macromolecular Complexes using Native Mass Spectrometry.- Hydrogen Deuterium Exchange Mass Spectrometry to Study Protein Complexes.- Structural Analysis of Protein Complexes by Cross-linking and Mass-spectrometry.- Global Characterization of Protein Complexes by Biochemical Purification-mass Spectrometry (BP/MS).- Proteomic Profiling of Integrin Adhesion Complex Assembly.- Dual-Color and 3D Super-Resolution Microscopy of Multi-protein Assemblies.- Correlative 3D Structured Illumination Microscopy and Single Molecular Localisation Microscopy for Imaging Cancer Invasion.- Observing the Assembly of Protein Complexes In Living Eukaryotic Cells In Super-resolution using refSOFI.- Detecting Purinosome Metabolon Formation with Fluorescence Microscopy.- Analysis of Bacterial pilus Assembly by Shearing and Immunofluorescence Microscopy.- Expression, Purification and Assembly of Archaellum Subcomplexes of Sulfolobus Acidocaldarius.- Unstable Protein Purification through the Formation of Stable Complexes.- Expressing Multi-subunit Complexes using biGBac.- Computational Modelling of Protein Complex Structure and Assembly.- Inferring and using Protein Quaternary Structure Information from Crystallographic.- Searching and Extracting Data from the EMBL-EBIComplex Portal.- Automated Computational Inference of Multi-protein Assemblies from Biochemical Co-Purification Data.- A Multi-scale Computational Model for Simulating the Kinetics of Protein Complex Assembly.- Flexible Protein-protein Docking with SwarmDock.- Protein-protein Docking using Evolutionary Information.- Modeling Structure and Dynamics of Protein Complexes with SAXS Profiles.- Modeling the Structure of Helical Assemblies with Experimental Constraints in.- Selecting Conformational Ensembles Using Residual Electron and Anomalous Density (READ).