Engineering Proteins for Stability and Efficient Folding

Abstract

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 Kinetic and Thermodynamic Aspects of Natural Proteins The Stability of Natural Proteins Different Kinds of “Stability” Thermodynamic Stability Kinetic Stability Folding Efficiency The Engineering Approach Consensus Strategies Principles Examples Structure‐based Engineering Entropic Stabilization Hydrophobic Core Packing Charge Interactions Hydrogen Bonding Disallowed Phi‐Psi Angles Local Secondary Structure Propensities Exposed Hydrophobic Side Chains Inter‐domain Interactions Case Study: Combining Consensus Design and Rational Engineering to Yield Antibodies with Favorable Biophysical Properties The Selection and Evolution Approach Principles Screening and Selection Technologies Available for Improving Biophysical Properties In Vitro Display Technologies Partial in Vitro Display Technologies In Vivo Selection Technologies Selection for Enhanced Biophysical Properties Selection for Solubility Selection for Protein Display Rates Selection on the Basis of Cellular Quality Control Selection for Increased Stability General Strategies Protein Destabilization Selections Based on Elevated Temperature Selections Based on Destabilizing Agents Selection for Proteolytic Stability Conclusions Acknowledgements

Structure-based Stability Engineering of the Mouse IgG1 Fab Fragment by Modifying Constant Domains

“…The stabilizing effect of the strengthened conformational rigidity of the proteins has been realized with proteins other than antibodies. 27 However, a rigid protein backbone, especially in the antibody domains, leads to decreased conformational flexibility, a property that may be deleterious with respect to their biological functions. Sequence comparison of C H 1 domains among mouse immunoglobulin subclasses, as is shown in Figure 2, indicates that C H 1 domains of IgA, IgG1, IgG3, IgG2a IgG2b, IgG2c, IgD and IgE have a quite high sequence variation in the loop comprising residues from 126 to 132, whereas the preceding residues Pro 123 and Leu 124 are highly conserved, as are the residues in the flanking β-strands.…”