Cell-free analysis of tail-anchor protein targeting to membranes.
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.
Henderson
Matthew P A
MP
Department of Biochemistry and Biomedical Sciences, McMaster University, Hamilton, Ont., Canada L8N 3Z5.
Billen
Lieven P
LP
Kim
Peter K
PK
Andrews
David W
DW
D002474:Cell-Free System / Q000737:chemistry*; D002845:Chromatography / Q000379:methods*; D008081:Liposomes / Q000737:chemistry / Q000378:metabolism; D008565:Membrane Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D017434:Protein Structure, Tertiary
D008081:Liposomes; D008565:Membrane Proteins