[Screening and identification interaction proteins of connexin 30].
OBJECTIVE
To explore interaction proteins affect functions of connexin 30 (Cx30) by screening and identification interaction proteins of Cx30.


METHODS
The fusion expression vecto of CX30-C-terminal functional domain-pGEX-4T-2-GST was constructed, and then, fusion protein and GST were purified. They were incubated with the proteins of the foetus brain tissue disruption to pull down interaction proteins. The interaction proteins were separated by SDS-PAGE. Differential straps were cut to enzymolysis to prepare for mass chromatographic analysis, and then to index and screen interaction proteins in NCBInr database. The interaction proteins were identified by immunolocalization.


RESULTS
The four interaction proteins of Cx30 were screened in the foetus brain tissue, as follow, Keratin 16, Camk2b, Tubulin beta-3 and alpha-tubulin. Cx30 was proved to coexist with Keratin 16 and Tubulin beta-3.


CONCLUSIONS
Keratin 16, Camk2b, Tubulin beta-3 and alpha-tubulin are the interaction proteins of Cx30. The interaction proteins affect the assembly, intracellular transport, and channel switch of Cx30.
He
Ding-hua
DH
Department of Otorhinolaryngology, Xiangya Hospital, Central South University, Changsha, China.
Feng
Yong
Y
Mei
Ling-yun
LY
He
Chu-feng
CF
D000075302:Connexin 30; D017630:Connexins / Q000235:genetics* / Q000378:metabolism; D005822:Genetic Vectors; D005982:Glutathione Transferase; D006801:Humans; D009152:Mutagenicity Tests; D025941:Protein Interaction Mapping; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism
D000075302:Connexin 30; D017630:Connexins; C491020:GJB6 protein, human; D011994:Recombinant Proteins; D005982:Glutathione Transferase