Identification of proteins interacted with Bat3 using tandem affinity purification.
- Author:
Wei WU
1
;
Qin-shan LI
1
;
Wei SONG
1
;
Shi-ying MIAO
1
;
Lin-fang WANG
1
Author Information
- Publication Type:Journal Article
- MeSH: Cell Line; Humans; Molecular Chaperones; isolation & purification; Protein Binding; Ubiquitins; isolation & purification
- From: Acta Academiae Medicinae Sinicae 2014;36(1):1-4
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo identify the specific protein interactions involved in Bat3-mediated apoptosis.
METHODSTandem affinity purification (TAP) was utilized to investigate Bat3-protein interactions, during which full-length human Bat3 fused with Strep2 and FLAG tag as a bait was used to screen the specific protein-protein interactions. The isolated proteins were identified with mass spectrometry.
RESULTSTAP studies showed that Ubl4A was identified as a Bat3-binding partner. Further investigation using co-immunoprecipitation confirmed that Bat3 was associated with Ubl4A.
CONCLUSIONTAP was successfully established and is suitable for isolating the binding partners of Bat3.
