Referenced in: none

Automatically associated channels: ClIC2 , Kv10.1

Title: 5,5'-Dithio-bis(2-nitrobenzoic acid) modification of cysteine improves the crystal quality of human chloride intracellular channel protein 2.

Authors: Wei Mi, Lanfen Li, Xiao-Dong Su

Journal, date & volume: Biochem. Biophys. Res. Commun., 2008 Apr 18 , 368, 919-22

PubMed link: http://www.ncbi.nlm.nih.gov/pubmed/18280248

Abstract
Structural studies of human chloride intracellular channel protein 2 (CLIC2) had been hampered by the problem of generating suitable crystals primarily due to the protein containing exposed cysteines. Several chemical reagents were used to react with the cysteines on CLIC2 in order to modify the redox state of the protein. We have obtained high quality crystals that diffracted to better than 2.5A at a home X-ray source by treating the protein with 5,5'-dithio-bis(2-nitrobenzoic acid) (DTNB). After solving the crystal structure of CLIC2, we found that the DTNB had reacted with the Cys(114), and made CLIC2 in a homogenous oxidized state. This study demonstrated that the DTNB modification drastically improved the crystallization of CLIC2, and it implied that this method may be useful for other proteins containing exposed cysteines in general.