Structural Analysis of Fibrous Proteins
Authors: Scheibel Thomas , Serpell Louise C. 
Published: March 2008

Originally published in: Protein Folding Handbook. Part II. Edited by Johannes Buchner and Thomas Kiefhaber. Copyright © 2005 Wiley‐VCH Verlag GmbH & Co. KGaA Weinheim. Print ISBN: 3‐527‐30784‐2

The sections in this article are

Introduction

Overview: Protein Fibers Formedin vivo

Amyloid Fibers

Silks

Collagens

Actin, Myosin, and Tropomyosin Filaments

Intermediate Filaments/Nuclear Lamina

Fibrinogen/Fibrin

Microtubules

Elastic Fibers

Flagella and Pili

Filamentary Structures in Rod‐like Viruses

Protein Fibers Used by Viruses and Bacteriophages to Bind to Their Hosts

Overview: Fiber Structures

Study of the Structure of β‐sheet‐containing Proteins

Amyloid

Paired Helical Filaments

β‐Silks

β‐Sheet‐containing Viral Fibers

α‐Helix‐containing Protein Fibers

Collagen

Tropomyosin

Intermediate Filaments

Protein Polymers Consisting of a Mixture of Secondary Structure

Tubulin

Actin and Myosin Filaments

Methods to Study Fiber Assembly

Circular Dichroism Measurements for Monitoring Structural Changes Upon Fiber Assembly

Theory of CD

Experimental Guide to Measure CD Spectra and Structural Transition Kinetics

Intrinsic Fluorescence Measurements to Analyze Structural Changes

Theory of Protein Fluorescence

Experimental Guide to Measure Trp Fluorescence

Covalent Fluorescent Labeling to Determine Structural Changes of Proteins with Environmentally Sensitive Fluorophores

Theory on Environmental Sensitivity of Fluorophores

Experimental Guide to Labeling Proteins With Fluorophores

1‐Anilino‐8‐Naphthalensulfonate (ANS) Binding to Investigate Fiber Assembly

Theory on UsingANSFluorescence for Detecting Conformational Changes in Proteins

Experimental Guide to UsingANSfor Monitoring Protein Fiber Assembly

Light Scattering to Monitor Particle Growth

Theory of Classical Light Scattering

Theory of Dynamic Light Scattering

Experimental Guide to Analyzing Fiber Assembly UsingDLS

Field‐flow Fractionation to Monitor Particle Growth

Theory ofFFF

Experimental Guide to UsingFFFfor Monitoring Fiber Assembly

Fiber Growth‐rate Analysis Using Surface Plasmon Resonance

Theory ofSPR

Experimental Guide to UsingSPRfor Fiber‐growth Analysis

Single‐fiber Growth Imaging Using Atomic Force Microscopy

Theory of Atomic Force Microscopy

Experimental Guide for UsingAFMto Investigate Fiber Growth

Dyes Specific for Detecting Amyloid Fibers

Theory on Congo Red and ThioflavinTBinding to Amyloid

Experimental Guide to Detecting Amyloid Fibers withCRand Thioflavin Binding

Methods to Study Fiber Morphology and Structure

Scanning Electron Microscopy for Examining the Low‐resolution Morphology of a Fiber Specimen

Theory ofSEM

Experimental Guide to Examining Fibers bySEM

Transmission Electron Microscopy for Examining Fiber Morphology and Structure

Theory ofTEM

Experimental Guide to Examining Fiber Samples byTEM

Cryo‐electron Microscopy for Examination of the Structure of Fibrous Proteins

Theory of Cryo‐electron Microscopy

Experimental Guide to Preparing Proteins for Cryo‐electron Microscopy

Structural Analysis from Electron Micrographs

Atomic Force Microscopy for Examining the Structure and Morphology of Fibrous Proteins

Experimental Guide for UsingAFMto Monitor Fiber Morphology

Use ofX‐ray Diffraction for Examining the Structure of Fibrous Proteins

Theory ofX‐Ray Fiber Diffraction

Experimental Guide toX‐Ray Fiber Diffraction

Fourier Transformed Infrared Spectroscopy

Theory ofFTIR

Experimental Guide to Determining Protein Conformation byFTIR

Conclusions

Acknowledgements