화학공학소재연구정보센터
Biochemical and Biophysical Research Communications, Vol.303, No.2, 475-482, 2003
One-step on-column affinity refolding purification and functional analysis of recombinant human VDACI
The outer mitochondrial membrane porin, voltage-dependent anion-selective channel (VDAC), is believed to play an important role in mediating mitochondria-dependent apoptosis. However, detailed structure-function studies of VDAC have been hindered by the difficulties to obtain a soluble, correctly folded, and fully active form of the recombinant VDAC and its mutant variants due to its transmembrane nature. Here we report a high-throughput one-step chromatographic procedure in purification of recombinant human VDAC1 (rhVDAC1) protein overexpressed in bacteria. The improved methodology could generate a large quantity of rhVDAC1 with correct folding in terms of the secondary structure, with full biological activities in mediating cytochrome c release and in interaction with Bcl-X-L. The method will significantly benefit genetic, biochemical, and structural studies of this critical channel protein. (C) 2003 Elsevier Science (USA). All rights reserved.