Research shows power of FRET-based approach for distinguishing among distinct states of proteins

Nov 30, 2009

In the December 2009 issue of the Journal of General Physiology, Moss et al. report a comprehensive investigation employing Förster resonance energy transfer (FRET) to study the {gamma}-amino acid (GABA) transporter GAT1, a member of the family that includes transporters for neurotransmitters dopamine (DAT), serotonin (SERT), norepinephrine (NET) and glycine (GlyT).

The investigators created a large panel of novel mouse GAT1 transporters tagged with cyan or yellow fluorescent proteins (CFP and YFP) and optimized their expression in . They determined the trafficking, subcellular localization, and oligomerization state of mGAT1 and correlated these features with transporter function.

One finding is that individual components of the FRET amplitude distribution reveal GAT1 dimers, high-order oligomers (likely tetramers), and oligomers associated via PDZ-mediated interactions with the actin cytoskeleton. Secondly, these details of the FRET amplitude distribution correlate with transporter function. Finally, the mGAT1 C-terminus PDZ-interacting domain is necessary for anchoring functional transporters to the actin cytoskeleton at the cell periphery; the corresponding FRET signal appears only in mGAT1 constructs with wild-type function. More generally, the results show the power of the FRET-based approach for distinguishing among distinct states of proteins.

More information: Moss, F.J., et al. 2009. J. Gen. Physiol. doi:10.1085/jgp.200910314

Source: Rockefeller University (news : web)

Explore further: Retinal regeneration in zebrafish (w/ Video)

add to favorites email to friend print save as pdf

Related Stories

Cocaine's effects on brain metabolism may contribute to abuse

Feb 18, 2008

Many studies on cocaine addiction - and attempts to block its addictiveness - have focused on dopamine transporters, proteins that reabsorb the brain's "reward" chemical once its signal is sent. Since cocaine blocks dopamine ...

Protein on 'speed' linked to ADHD

Jul 08, 2008

A genetic change in the dopamine transporter – one of the brain's dopamine-handling proteins – makes it behave as if amphetamine is present and "run backward," Vanderbilt University Medical Center investigators report ...

Recommended for you

Diet affects men's and women's gut microbes differently

Jul 29, 2014

The microbes living in the guts of males and females react differently to diet, even when the diets are identical, according to a study by scientists from The University of Texas at Austin and six other institutions published ...

User comments : 0