Improved identification of SUMO attachment sites using C-terminal SUMO mutants and tailored protease digestion strategies.
J Proteome Res
; 5(4): 761-70, 2006 Apr.
Article
em En
| MEDLINE
| ID: mdl-16602682
ABSTRACT
A growing number of biological processes have been found to be regulated by the covalent attachment of the ubiquitin-like protein SUMO to key cellular targets. A critical step in the process of analyzing the role of SUMO in regulating the activity of these proteins is the identification of the lysine residues that are targeted by this modification. Unfortunately, current methods aimed at mapping these attachment-sites are laborious and often ineffective. We report here the development of a platform that combines the use of different C-terminal SUMO mutants with different protease digestion strategies to enable the rapid and efficient identification of SUMO attachment sites. We successfully apply this approach to several model SUMO substrates as well as to a mixture of SUMO conjugates purified from Saccharomyces cerevisiae. Although we specifically employ this strategy for the identification of SUMO attachment sites in yeast, this general approach can easily be adapted to map the sites of conjugation for other ubiquitin-like proteins from a wide range of organisms.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Proteínas Fúngicas
/
Proteínas Modificadoras Pequenas Relacionadas à Ubiquitina
/
Mutação
Tipo de estudo:
Diagnostic_studies
/
Prognostic_studies
Idioma:
En
Revista:
J Proteome Res
Assunto da revista:
BIOQUIMICA
Ano de publicação:
2006
Tipo de documento:
Article
País de afiliação:
Estados Unidos