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Bulletin of the Korean Chemical Society (BKCS)

ISSN 0253-2964(Print)
ISSN 1229-5949(Online)
Volume 28, Number 4
BKCSDE 28(4)
April 20, 2007 

The Binding of Human CLIC1 with SEDL and Its Characterization in vitro
Jeong Soon Park, Kyoung Mi Lee, Mi Suk Jeong, Gyoung-Ean Jin, Se Bok Jang*
Subcloning, Overexpression, Purification, Characterization, Chloride intracellular channel (CLIC1)
Full-length chloride intracellular channel protein 1 (CLIC1) is a member of the family of proteins related to bovine chloride intracellular channel p64. Mutations in the SEDL gene cause spondyloepiphyseal dysplasia tarda (SEDT), a rare X-linked chondrodysplasia. The link between the intracellular chloride channels and SEDL is an important step toward understanding their functional interplay. In the present study, CLIC1 protein was subcloned into the pGEX-KG vector and overexpressed in XL-1 blue cells. We developed a large-scale expression system composed of glutathione S-transferase (GST) fused with a 240-amino-acid CLIC1 protein in Escherichia coli. The soluble CLIC1 protein was successfully purified to homogeneity, and its purity, identity, activity and conformation were determined using SDS-PAGE, MALDI-MS, biophotometer and circular dichroism spectroscopic studies. The binding of both CLIC1 and SEDL proteins in vitro was detected by BIAcore biosensor and fluorescence measurements.
574 - 580
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