Biology Reference
In-Depth Information
Table 18.2
Composition of Brain Polar Lipid
Component
w/w%
PC
12.6
PE
33.1
PI
4.1
PS
18.5
PA
0.8
Unknown
30.9
Details from Avanti Polar lipids. PC, phosphatidylcholine; PE,
phosphatidylethanolamine; PI, phosphatidylinositol; PS,
phosphatidylserine; PA, phosphatidic acid. At least some of the
“unknown” is cholesterol (our observations).
Polar Lipids) that has the composition shown in
Table 18.2
. BPL is the best mimic of
the endogenous environment of NTS1—it is unsurprising that NTS1 dimerizes effi-
ciently in this lipid mixture. To determine the contribution of the various lipids to
dimerization, we have performed FRET experiments in liposomes of defined com-
position. This demonstrates that presence of POPE and cholesterol strongly influ-
ences dimerization.
SUMMARY
Here, we have given details of the methods used to reconstitute a GPCR with a view
to studying dimerization. The approaches could well be generic, but firstly, only few
GPCRs are expressed either folded or in inclusion bodies and require refolding. Sec-
ondly, there is still too little information available at the moment to make general-
izations about detergent suitability for GPCRs—we are likely to require more
detergents or methods of GPCR solubilization, and some generalizations may be-
come evident as more biophysical studies are performed that require reconstituted
receptor.
References
