Cell-free analysis of tail-anchor protein targeting to membranes.

Matthew P A Henderson, Lieven P Billen, Peter K Kim, David W Andrews

Department of Biochemistry and Biomedical Sciences, McMaster University, Hamilton, Ont., Canada L8N 3Z5.

Methods (San Diego, Calif.) 2007 Apr

filter terms:

We describe cell-free strategies for analysis of the biogenesis of membrane proteins. Special emphasis is placed on the unique challenges presented by tail-anchor proteins for the analysis of: targeting specificity, binding to membranes such as endoplasmic reticulum, mitochondria and pure lipid membranes, protein topology in the membrane and the dependence of various steps on additional proteins. Tail-anchor proteins such as cytochrome b(5), Bcl-2 and Bcl-X(L) are used to address the suitability of both cell-free techniques based on in vitro transcription translation systems and the use of purified recombinant protein and pure lipid membranes. Together these cell-free systems permit detailed characterization of membrane protein biogenesis.

Citation

Matthew P A Henderson, Lieven P Billen, Peter K Kim, David W Andrews. Cell-free analysis of tail-anchor protein targeting to membranes. Methods (San Diego, Calif.). 2007 Apr;41(4):427-38