title: Interplay between Protein Thermal Flexibility and Kinetic Stability.
abstract: Kinetic stability is a key parameter to comprehend protein behavior and it plays a central role to understand how evolution has reached the balance between function and stability in cell-relevant timescales. Using an approach that includes simulations, protein engineering, and calorimetry, we show that there is a clear correlation between kinetic stability determined by differential scanning calorimetry and protein thermal flexibility obtained from a novel method based on temperature-induced unfolding molecular dynamics simulations. Thermal flexibility quantitatively measures the increment of the conformational space available to the protein when energy in provided. The (β/α)8barrel fold of two closely related by evolution triosephosphate isomerases from two trypanosomes are used as model systems. The kinetic stability-thermal flexibility correlation has predictive power for the studied proteins, suggesting that the strategy and methodology discussed here might be applied to other proteins in biotechnological developments, evolutionary studies, and the design of protein based therapeutics.
authors.affiliation: Laboratorio de Biofisicoquímica, Departamento de Fisicoquímica, Facultad de Química, Universidad Nacional Autónoma de México, México City 04510, México.
authors.forename: Andrea G
authors.initials: AG
authors.lastname: Quezada
authors.affiliation: Laboratorio de Biofisicoquímica, Departamento de Fisicoquímica, Facultad de Química, Universidad Nacional Autónoma de México, México City 04510, México.
authors.forename: A Jessica
authors.initials: AJ
authors.lastname: Díaz-Salazar
authors.affiliation: Departamento de Bioquímica y Biología Estructural, Instituto de Fisiología Celular, Universidad Nacional Autónoma de México, Mexico City 04510, México.
authors.forename: Nallely
authors.initials: N
authors.lastname: Cabrera
authors.affiliation: Departamento de Bioquímica y Biología Estructural, Instituto de Fisiología Celular, Universidad Nacional Autónoma de México, Mexico City 04510, México.
authors.forename: Ruy
authors.initials: R
authors.lastname: Pérez-Montfort
authors.affiliation: Soft Matter and Molecular Biophysics Group, Department of Applied Physics, University of Santiago de Compostela, Santiago de Compostela 15782, Spain. Electronic address: angel.pineiro@usc.es.
authors.forename: Ángel
authors.initials: Á
authors.lastname: Piñeiro
authors.affiliation: Laboratorio de Biofisicoquímica, Departamento de Fisicoquímica, Facultad de Química, Universidad Nacional Autónoma de México, México City 04510, México. Electronic address: costasmi@unam.mx.
authors.forename: Miguel
authors.initials: M
authors.lastname: Costas
mesh_terms: D002152:Calorimetry, Differential Scanning; D007700:Kinetics; D008968:Molecular Conformation; D011489:Protein Denaturation; D015202:Protein Engineering; D055550:Protein Stability; D017433:Protein Structure, Secondary; D058767:Protein Unfolding; D013816:Thermodynamics; D014305:Triose-Phosphate Isomerase / Q000737:chemistry*; D014345:Trypanosoma / Q000737:chemistry / Q000201:enzymology*
chemical_list: D014305:Triose-Phosphate Isomerase
keywords: DSC; TIM barrel; activation energy; calorimetry; chimeras; kinetic stability; protein flexibility; triosephosphate isomerase; unfolding MD; unfolding cooperativity