Rational stabilization of complex proteins: A divide and combine approach
Resumen: Increasing the thermostability of proteins is often crucial for their successful use as analytic, synthetic or therapeutic tools. Most rational thermostabilization strategies were developed on small two-state proteins and, unsurprisingly, they tend to fail when applied to the much more abundant, larger, non-fully cooperative proteins. We show that the key to stabilize the latter is to know the regions of lower stability. To prove it, we have engineered apoflavodoxin, a non-fully cooperative protein on which previous thermostabilizing attempts had failed. We use a step-wise combination of structure-based, rationally-designed, stabilizing mutations confined to the less stable structural region, and obtain variants that, according to their van't Hoff to calorimetric enthalpy ratios, exhibit fully-cooperative thermal unfolding with a melting temperature of 75°C, 32 degrees above the lower melting temperature of the non-cooperative wild type protein. The ideas introduced here may also be useful for the thermostabilization of complex proteins through formulation or using specific stabilizing ligands (e.g. pharmacological chaperones).
Idioma: Inglés
DOI: 10.1038/srep09129
Año: 2015
Publicado en: Scientific Reports 5 (2015), 9129 [11 pp.]
ISSN: 2045-2322

Factor impacto JCR: 5.228 (2015)
Categ. JCR: MULTIDISCIPLINARY SCIENCES rank: 7 / 62 = 0.113 (2015) - Q1 - T1
Factor impacto SCIMAGO: 2.034 - Multidisciplinary (Q1)

Financiación: info:eu-repo/grantAgreement/ES/DGA/B89
Financiación: info:eu-repo/grantAgreement/ES/MICINN/BFU2010-16297
Financiación: info:eu-repo/grantAgreement/ES/MINECO/BFU2013-47064-P
Tipo y forma: Article (Published version)
Área (Departamento): Área Química Analítica (Dpto. Química Analítica)
Área (Departamento): Área Bioquímica y Biolog.Mole. (Dpto. Bioq.Biolog.Mol. Celular)

Creative Commons You must give appropriate credit, provide a link to the license, and indicate if changes were made. You may do so in any reasonable manner, but not in any way that suggests the licensor endorses you or your use.

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