A proximity-tagging system to identify membrane protein-protein interactions.
Nat Methods
; 15(9): 715-722, 2018 09.
Article
en En
| MEDLINE
| ID: mdl-30104635
ABSTRACT
The communication between cells and between cellular organelles is often controlled by the interaction of membrane proteins. Although many methods for the detection of protein-protein interactions (PPIs) exist, membrane PPIs remain difficult to detect. Here we developed a proximity-based tagging system, PUP-IT (pupylation-based interaction tagging), to identify membrane protein interactions. In this approach, a small protein tag, Pup, is applied to proteins that interact with a PafA-fused bait, enabling transient and weak interactions to be enriched and detected by mass spectrometry. Pup does not diffuse from the enzyme, which allows high-specificity labeling. We applied this approach to CD28, a critical costimulatory receptor for T lymphocyte activation, and identified known CD28 binding partners and multiple potential interacting proteins. In addition, we demonstrated that this method can identify the interaction between a cell surface receptor and its ligand.
Texto completo:
1
Banco de datos:
MEDLINE
Asunto principal:
Mapas de Interacción de Proteínas
/
Proteínas de la Membrana
Límite:
Humans
Idioma:
En
Año:
2018
Tipo del documento:
Article