Engineering and structural characterization of a linear polyubiquitin-specific antibody.
J Mol Biol
; 418(3-4): 134-44, 2012 May 04.
Article
en En
| MEDLINE
| ID: mdl-22227388
ABSTRACT
Polyubiquitination is an essential posttranslational modification that plays critical roles in cellular signaling. PolyUb (polyubiquitin) chains are formed by linking the carboxyl-terminus of one Ub (ubiquitin) subunit to either a lysine residue or the amino-terminus of an adjacent Ub. Linkage through the amino-terminus results in linear polyubiquitination that has recently been demonstrated to be a key step in nuclear factor κB activation; however, tools to study linear chains have been lacking. We therefore engineered a linear-linkage-specific antibody that is functional in Western blot, immunoprecipitation, and immunofluorescence applications. A crystal structure of the linear-linkage-specific antibody Fab fragment in complex with linear diubiquitin provides molecular insight into the nature of linear chain specificity. We use the antibody to demonstrate that linear polyUb is up-regulated upon tumor necrosis factor α stimulation of cells, consistent with a critical role in nuclear factor κB signaling. This antibody provides an essential tool for further investigation of the function of linear chains.
Texto completo:
1
Colección:
01-internacional
Banco de datos:
MEDLINE
Asunto principal:
Fragmentos Fab de Inmunoglobulinas
/
Poliubiquitina
/
Anticuerpos
Tipo de estudio:
Prognostic_studies
Idioma:
En
Revista:
J Mol Biol
Año:
2012
Tipo del documento:
Article
País de afiliación:
Estados Unidos