Localization of SUMO-modified Proteins Using Fluorescent Sumo-trapping Proteins.
J Vis Exp
; (146)2019 04 27.
Article
en En
| MEDLINE
| ID: mdl-31081817
Here we are presenting a novel method to study the sumoylation of proteins and their sub-cellular localization in mammalian cells and nematode oocytes. This method utilizes a recombinant modified SUMO-trapping protein fragment, kmUTAG, derived from the Ulp1 SUMO protease of the stress-tolerant budding yeast Kluyveromyces marxianus. We have adapted the properties of the kmUTAG for the purpose of studying sumoylation in a variety of model systems without the use of antibodies. For the study of SUMO, KmUTAG has several advantages when compared to antibody-based approaches. This stress-tolerant SUMO-trapping reagent is produced recombinantly, it recognizes native SUMO isoforms from many species, and unlike commercially available antibodies it shows reduced affinity for free, unconjugated SUMO. Representative results shown here include the localization of SUMO conjugates in mammalian tissue culture cells and nematode oocytes.
Texto completo:
1
Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Proteínas Modificadoras Pequeñas Relacionadas con Ubiquitina
Límite:
Animals
Idioma:
En
Revista:
J Vis Exp
Año:
2019
Tipo del documento:
Article
Pais de publicación:
Estados Unidos