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: Growing a blood vessel in a week

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

Growing a blood vessel in a week

Oct 24, 2014

The technology for creating new tissues from stem cells has taken a giant leap forward. Three tablespoons of blood are all that is needed to grow a brand new blood vessel in just seven days. This is shown ...

Testing time for stem cells

Oct 24, 2014

DefiniGEN is one of the first commercial opportunities to arise from Cambridge's expertise in stem cell research. Here, we look at some of the fundamental research that enables it to supply liver and pancreatic ...

Team finds key signaling pathway in cause of preeclampsia

Oct 23, 2014

A team of researchers led by a Wayne State University School of Medicine associate professor of obstetrics and gynecology has published findings that provide novel insight into the cause of preeclampsia, the leading cause ...

Rapid test to diagnose severe sepsis

Oct 23, 2014

A new test, developed by University of British Columbia researchers, could help physicians predict within an hour if a patient will develop severe sepsis so they can begin treatment immediately.

User comments : 0