Explore the Structural Domains of CENP-E Protein Interacting with Mps1 Protein by FRET Method
- VernacularTitle:用FRET方法研究与Mps1蛋白有相互作用的CENP-E蛋白结构域
- Author:
Zi-Jie LIU
;
Ya-Guang WENG
;
Su-Yan LI
;
Qiong SI
;
Yan CAI
;
Bin LIU
;
Yan ZHANG
;
Chen YAN
;
- Publication Type:Journal Article
- Keywords:
CENP-E Mps1 Protein-protein interaction Fluorescence resonance energy transfer
- From:
China Biotechnology
2006;0(04):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To explore the structural domains of the CENP-E protein that interact with Mps1 protein.Methods: Two recombinant vectors named pEGFP-CENPE2(containing 674-1085 amino acids of CENP-E protein) and pEGFP-CENPE 3(containing 1200~2134 amino acids of CENP-E protein) were transfected into human embryo kidney 293(HEK293) cells respectively.The respective energy transfer efficiency(Ef) between either EGFP-CENPE2 and Mps1,or EGFP-CENPE3 and Mps1 were detected by FRET through selective photobleaching of the acceptors.Results: Both recombinant proteins expressed in HEK293 cells transfected by the recombinant plasmids were found to co-localize with the Mps1 protein as confirmed by confocal microscopy.The Ef between EGFP-CENPE3 and Mps1 protein was [(12.63?0.48)%,n=30] and that between EGFP-CENPE3 and Mps1 protein was [(3.17?0.21)%,n=30] as revealed by the results from FRET,the result of FRET was confirmed by co-Immunoprecipitate(CO-IP) method.When compared with that between the control and Mps1,the Ef between EGFP-CENPE3 and Mps1 was significantly higher(p
