Article

Direct Assessment of the Effect of the Gly380Arg Achondroplasia Mutation on FGFR3 Dimerization Using Quantitative Imaging FRET

Department of Materials Science and Engineering, Johns Hopkins University, Baltimore, Maryland, United States of America.
PLoS ONE (Impact Factor: 3.23). 10/2012; 7(10):e46678. DOI: 10.1371/journal.pone.0046678
Source: PubMed

ABSTRACT

The Gly380Arg mutation in FGFR3 is the genetic cause for achondroplasia (ACH), the most common form of human dwarfism. The mutation has been proposed to increase FGFR3 dimerization, but the dimerization propensities of wild-type and mutant FGFR3 have not been compared. Here we use quantitative imaging FRET to characterize the dimerization of wild-type FGFR3 and the ACH mutant in plasma membrane-derived vesicles from HEK293T cells. We demonstrate a small, but statistically significant increase in FGFR3 dimerization due to the ACH mutation. The data are consistent with the idea that the ACH mutation causes a structural change which affects both the stability and the activity of FGFR3 dimers in the absence of ligand.

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    • "These vesicles are derived from the native cellular membrane, and are thus more native-like than vesicles made of synthetic lipids. They are increasingly used in studies of lipid–lipid, lipid–protein and protein–protein interactions, and have already yielded new knowledge about lipid domains and receptor interactions in the membrane [11] [12] [13] [14] [15]. Often, however, they are not well characterized in terms of their lipid and protein content. "
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    • "The plasma membrane derived vesicles lack a transmembrane potential and a cytoskeleton, and are not a perfect mimic of the plasma membrane. Yet, plasma membrane derived vesicles are the only model system that has allowed us thus far to characterize the interaction strength between glycosylated membrane proteins in quantitative terms [16] [23]. "
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