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~


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