Miniaturised interaction proteomics on a microfluidic platform with ultra-low input requirements.
Nat Commun
; 10(1): 1525, 2019 04 04.
Article
en En
| MEDLINE
| ID: mdl-30948724
ABSTRACT
Essentially all cellular processes are orchestrated by protein-protein interactions (PPIs). In recent years, affinity purification coupled to mass spectrometry (AP-MS) has been the preferred method to identify cellular PPIs. Here we present a microfluidic-based AP-MS workflow, called on-chip AP-MS, to identify PPIs using minute amounts of input material. By using this automated platform we purify the human Cohesin, CCC and Mediator complexes from as little as 4 micrograms of input lysate, representing a 50â100-fold downscaling compared to regular microcentrifuge tube-based protocols. We show that our platform can be used to affinity purify tagged baits as well as native cellular proteins and their interaction partners. As such, our method holds great promise for future biological and clinical AP-MS applications in which sample amounts are limited.
Texto completo:
1
Bases de datos:
MEDLINE
Asunto principal:
Microfluídica
/
Mapas de Interacción de Proteínas
Tipo de estudio:
Guideline
Límite:
Humans
Idioma:
En
Revista:
Nat Commun
Asunto de la revista:
BIOLOGIA
/
CIENCIA
Año:
2019
Tipo del documento:
Article
País de afiliación:
Países Bajos