In addition to Bert remarks, you can read this paper from the positive
inside rule instigators.
Seppälä S, Slusky JS, Lloris-Garcerá P, Rapp M, von Heijne G. Control of
membrane protein topology by a single C-terminal residue. Science. 2010
Jun 25;328(5986):1698-700. Epub 2010 May 27. PubMed PMID: 20508091.
with the cited literature
Daniel
Le 04/03/2011 17:31, Justin Hall a écrit :
Dear Community,
In trying to trouble shoot an experiment I have become interested in the
cellular process that regulates the insertion and proper orientation of
membrane proteins. I am looking for references for how a GPCR is
correctly oriented during expression (i.e. the extra cellular domain
ends up extra cellularly oriented instead of a 50/50 mix in and out), my
intuition is that there must be an N-terminal sequence that directs this
process, but I am having no luck finding information on what this
sequence is for GPCRs, what players are involved or how orientation is
thought to be controlled. Any suggestions?
This is all spurred by my wanting to use phage display with a protein
that binds to the intracellular side of a GPCR, but of course that is
the hard side to present to the outside of a cell so I need to figure
out how to flip these guys around. I have thought about adding a new TM
helix before TM1 (or removing TM1) to flip these guys, but was hoping
there might be another way around that doesn't involve such massive
architectural rearrangement such as simply clipping the N-terminal
sequence responsible for proper orientation (if such a thing exists).
Cheers~
~Justin
