GMMA Can Stabilize Proteins Across Different Functional Constraints.
J Mol Biol
; 436(11): 168586, 2024 Jun 01.
Article
en En
| MEDLINE
| ID: mdl-38663544
ABSTRACT
Stabilizing proteins without otherwise hampering their function is a central task in protein engineering and design. PYR1 is a plant hormone receptor that has been engineered to bind diverse small molecule ligands. We sought a set of generalized mutations that would provide stability without affecting functionality for PYR1 variants with diverse ligand-binding capabilities. To do this we used a global multi-mutant analysis (GMMA) approach, which can identify substitutions that have stabilizing effects and do not lower function. GMMA has the added benefit of finding substitutions that are stabilizing in different sequence contexts and we hypothesized that applying GMMA to PYR1 with different functionalities would identify this set of generalized mutations. Indeed, conducting FACS and deep sequencing of libraries for PYR1 variants with two different functionalities and applying a GMMA analysis identified 5 substitutions that, when inserted into four PYR1 variants that each bind a unique ligand, provided an increase of 2-6 °C in thermal inactivation temperature and no decrease in functionality.
Palabras clave
Texto completo:
1
Base de datos:
MEDLINE
Asunto principal:
Reguladores del Crecimiento de las Plantas
/
Proteínas de Plantas
/
Análisis Mutacional de ADN
/
Ingeniería de Proteínas
/
Receptores de Superficie Celular
/
Estabilidad Proteica
Idioma:
En
Revista:
J Mol Biol
Año:
2024
Tipo del documento:
Article
País de afiliación:
Estados Unidos