Tuning Enzyme Thermostability via Computationally Guided Covalent Stapling and Structural Basis of Enhanced Stabilization
Jacob A. Iannuzzelli, J.P. Bacik, Eric J. Moore, Zhuofan Shen, Ellen M. Irving, David A. Vargas, Sagar D. Khare, Nozomi Ando, Rudi Fasan
Abstract
= +30 °C) were obtained while maintaining high levels of catalytic activity and stereoselectivity. Crystallographic analyses of singly and doubly stapled variants provide key insights into the structural basis for stabilization, which includes both direct interactions of the staples with protein residues and indirect interactions through adjacent residues involved in heme binding. This work expands the toolbox of protein stapling strategies available for protein stabilization.
Topics & Concepts
ThermostabilityChemistryProtein engineeringHemeProtein designProtein structureAmino acidBiocatalysisCysteineCombinatorial chemistryEnzymeStereochemistryBiochemistryCatalysisReaction mechanismProtein Structure and DynamicsEnzyme Catalysis and ImmobilizationEnzyme Structure and Function