Instrumental Analysis of Intrinsically Disordered Proteins Assessing Structure and Conformation

by Uversky, Vladimir; Longhi, Sonia

• ISBN13: 9780470343418
• ISBN10: 0470343419
• Edition: 1st
• Format: Hardcover
• Copyright: 2010-04-12
• Publisher: Wiley

Summary

This text offers the first systematic compilation of various techniques and methodologies used in the analysis of intrinsically disordered proteins (IDPs). Its coverage addresses one of the hottest yet least documented areas in protein biophysics today, summarizing recent achievements in the methods for structural characterization of intrinsically disordered proteins. The text also addresses structural biology and protein degradation, along with a myriad of spectroscopic techniques that can be used to elucidate protein structure: NMR and EPR spectroscopies, FTIR, circular dichroism, fluorescence spectroscopy, vibrational methods, and single-molecule analysis.

Author Biography

Vladimir N. Uversky, PhD, is a Senior Research Professor at the Indiana University School of Medicine. He obtained his academic degrees from Moscow Institute of Physics and Technology (PhD) and the Institute of Theoretical and Experimental Biophysics, Russian Academy of Sciences. Professor Uversky has authored over 300 scientific publications and edited several books and book series on protein structure, function, folding, and misfolding. He is also an editor of several scientific journals. Sonia Longhi, PhD, is a Director of Research at the Center for the National Scientific Research (CNRS), heading the Structural Disorder and Molecular Recognition group within the AFMB laboratory. She obtained her PhD from the Universit degli Studi of Milan in 1993. She has authored more than sixty scientific publications and edited a book on measles virus nucleoprotein.

Table of Contents

Preface	p. ix
Introduction to the Wiley Series on Protein and Peptide Science	p. xvii
List of Contributors	p. xix
List of Abbreviations	p. xxv
Assessing IDPs in the Living Cell	p. 1
IDPs and Protein Degradation in The Cell	p. 3
The Structural Biology of IDPs Inside Cells	p. 37
Spectroscopic Techniques	p. 59
Nuclear Magnetic Resonance Spectroscopy Applied to (Intrinsically) Disordered Proteins	p. 61
Atomic-Level Characterization of Disordered Protein Ensembles Using NMR Residual Dipolar Couplings	p. 89
Determining Structural Ensembles for Intrinsically Disordered Proteins	p. 107
Site-Directed Spin Labeling EPR Spectroscopy	p. 131
The Structure of Unfolded Peptides and Proteins Explored by Vibrational Spectroscopy	p. 171
Intrinsically Disordered Proteins and Induced Folding Studied by Fourier Transform Infrared Spectroscopy	p. 225
Genetically Engineered Polypeptides as a Model of Intrinsically Disordered Fibrillogenic Proteins: Deep UV Resonance Raman Spectroscopic Study	p. 253
Circular Dichroism of Intrinsically Disordered Proteins	p. 303
Fluorescence Spectroscopy of Intrinsically Disordered Proteins	p. 323
Hydration of Intrinsically Disordered Proteins From Wide-Line NMR	p. 345
Single-Molecule Techniques	p. 369
Single-Molecule Spectroscopy of Unfolded Proteins	p. 371
Monitoring the Conformational Equilibria of Monomeric Intrinsically Disordered Proteins by Single-Molecule Force Spectroscopy	p. 391
Methods to Assess Protein Size and Shape	p. 431
Analytical Ultracentrifugation, a Useful Tool to Probe Intrinsically Disordered Proteins	p. 433
Structural Insights into Intrinsically Disordered Proteins by Small-Angle X-Ray Scattering	p. 451
Dynamic and Static Light Scattering	p. 477
Analyzing Intrinsically Disordered Proteins by Size Exclusion Chromatography	p. 525
Conformational Stability	p. 545
Conformational Behavior of Intrinsically Disordered Proteins: Effects of Strong Denaturants, Temperature, PH, Counterions, and Macromolecular Crowding	p. 547
Detecting Disordered Regions in Proteins by Limited Proteolysis	p. 569
Mass Spectrometry	p. 627
Mass Spectrometry Tools for the Investigation of Structural Disorder and Conformational Transitions in Proteins	p. 629
Expression and Purification of IDPs	p. 653
Recombinant Production of Intrinsically Disordered Proteins for Biophysical and Structural Characterization	p. 655
Large-Scale Identification of Intrinsically Disordered Proteins	p. 671
Purification of Intrinsically Disordered Proteins	p. 695
Index	p. 705
Table of Contents provided by Ingram. All Rights Reserved.