Work

Protein thermostabilization with Protposer: Pushing the stability limits and folding reversibility of a highly-stabilized apoflavodoxin

Year: 2025

Type: article

Abstract: of 10 M apoflavodoxin is nearly 9 °C higher, while the simplified 3 M and 4 M mutants, showing improved refolding properties, display increases of 6/7.5 °C, respectively. The thermostabilizing effects of individual mutations are close to additive and translate into a large increase in conformational stability at 25.0 °C. Comparison of the x-ray structures of progressively stabilized WT, 6 M and 10 M flavodoxins reveals a concomitant mild trend toward shorter hydrogen bonds, reduced internal cavity volumes and narrower tunnels. Overall, these conformational changes are minor, and a functional assay confirms the mutants also preserve their catalytic activity. These findings demonstrate that even highly stable proteins can be further stabilized through rational design using a simple computational tool that automatically analyses PDB files and identifies stabilizing mutations.

Source: International Journal of Biological Macromolecules

DOI: 10.1016/j.ijbiomac.2025.148333

Authors: Antonio Hidalgo-Toledo, Darío Bazco, Víctor Correa-Pérez, Marta Martínez-Júlvez, Javier Sancho

Institutions: Universidad de Zaragoza, Instituto de Biocomputación y Física de Sistemas Complejos, Instituto de Investigación Sanitaria Aragón

Language: English

Topic: Protein Structure and Dynamics

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: hybrid

Estimated APC paid: $4,390