Rational stabilization of complex proteins: A divide and combine approach

Llamazares, Emilio; Sancho Sanz, Javier; Bueno Fernandez, Marta; Clemente Arellano, Isabel; Velázquez Campoy, Adrián

doi:10.1038/srep09129

Rational stabilization of complex proteins: A divide and combine approach

Llamazares, Emilio ; Clemente Arellano, Isabel (Universidad de Zaragoza) ; Bueno Fernandez, Marta ; Velázquez Campoy, Adrián (Universidad de Zaragoza) ; Sancho Sanz, Javier (Universidad de Zaragoza)

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 / 60 = 0.117 (2015) - Q1 - T1
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): Química Analítica (Departamento de Química Analítica)
Área (Departamento): Bioquímica y Biología Molecular (Departamento de Bioquímica y Biología Molecular y Celular)
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Este artículo se encuentra en las siguientes colecciones:
articulos > articulos-por-area > bioquimica_y_biologia_molecular
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Rational stabilization of complex proteins: A divide and combine approach