BINA alone had no effect on FRET histograms of SNAP-labeled, full-length mGlu2 in LMNG-CHS-GDN micelles. In immobilized SNAP-mGluR2-TMD molecules, BINA application did not change the proportion of higher order complexes assessed by photobleaching step distributions. In the VFT closure sensor, adding BINA had no significant additional effect compared with glutamate alone. In IGEPAL-solubilized human mGlu2 receptors already totally stabilized in the active VFT state by Glu, adding BINA produced no further effect on VFT activation. [1,2,3]
Literature-reported observations and conditions; not Adooq-tested protocols or general treatment recommendations.
FRET histograms of micellar mGlu2
Cell-based study · SNAP-labeled, full-length mGlu2 in LMNG-CHS-GDN micelles [1]
SNAP-labeled, full-length mGlu2 in LMNG-CHS-GDN micelles was treated with BINA at 10 µM. Compared with the absence of ligands, BINA alone had no effect on FRET histograms.
Literature-Reported Treatment Conditions
- Treatment concentration
- 10 µM
- Vehicle
- acquisition buffer
- Reported limits
- Not stated in the sentences used: route, schedule.
Methods and source locations
Compound reported in the paper: BINA
Evidence for BINA as the hydrochloride form reported in the paper, not the free base/neutral form supplied as A11126; no salt equivalence, dose conversion or lot validation.
Results > Allosteric modulation through the 7TM is required to stabilize the fully active VFT state (S35, S37, S42); Methods > Transfection and labeling (S121); Methods > smFRET measurements (S163, S165); Fig. 3 legend (S207)
FRET histograms: FRET histograms; comparator: in the absence of ligands — Results > Allosteric modulation through the 7TM is required to stabilize the fully active VFT state (S35, S42)
Photobleaching distributions in immobilized receptor molecules
Cell-based study · immobilized SNAP-mGluR2-TMD molecules [2]
BINA was applied at 1 µM to immobilized SNAP-mGluR2-TMD molecules. Photobleaching distributions showed ~40–45% two-step bleaching events, with no change in the proportion of higher order complexes with >3 bleaching steps (<10%).
Literature-Reported Treatment Conditions
- Treatment concentration
- 1 µM
- Method
- application
- Reported limits
- Not stated in the sentences used: schedule, vehicle.
Methods and source locations
Compound reported in the paper: BINA
Literature evidence for the named compound in the stated model; no Adooq product-use or lot-performance claim.
Results > mGluR TMDs form dimers of variable propensity and assembly of mGluR2-TMD dimers is insensitive to allosteric drugs (S67); Materials and methods > Single-molecule pulldown and subunit counting (S200, S201); Figure 2. legend (S285, S286); Figure 2—figure supplement 3. legend (S308)
photobleaching step distributions: photobleaching step distributions; comparator: un-liganded SNAP-mGluR2-TMD — Results > mGluR TMDs form dimers of variable propensity and assembly of mGluR2-TMD dimers is insensitive to allosteric drugs (S67)
VFT closure with glutamate
Cell-based study · VFT closure sensor [3]
A VFT closure sensor measured the fraction of molecules in the closed state with BINA at 1 μM and glutamate at 10 mM. Compared with glutamate alone, BINA had no significant additional effect.
Literature-Reported Treatment Conditions
- Treatment concentration
- 1 μM
- Co-treatment
- Other interventions in this design: glutamate 10 mM. Compound effect isolated by the comparison: glutamate alone.
- Reported limits
- Not stated in the sentences used: route, schedule, vehicle.
Methods and source locations
Compound reported in the paper: BINA
Literature evidence for the named compound in the stated model; no Adooq product-use or lot-performance claim.
RESULTS > Full agonists induce maximal VFT closure independently of allosteric modulation (S70, S76); Fig. 3. legend (S260, S264)
Measured fraction of molecules in the closed state: Measured fraction of molecules in the closed state; comparator: 10 mM glutamate — RESULTS > Full agonists induce maximal VFT closure independently of allosteric modulation (S76); Fig. 3. legend (S264)
BINA addition and VFT activation
Cell-based study · human mGlu2 receptors [1]
In receptors solubilized with IGEPAL, Glu was sufficient to totally stabilize the receptors in the active VFT state. No further effect on VFT activation was observed upon the addition of BINA.
Literature-Reported Treatment Conditions
- Treatment concentration
- 10 µM
- Vehicle
- acquisition buffer
- Co-treatment
- Other interventions in this design: Glu 10 mM. Compound effect isolated by the comparison: Glu was sufficient to totally stabilize the receptors in the active VFT state.
- Reported limits
- Not stated in the sentences used: route, schedule.
Methods and source locations
Compound reported in the paper: BINA
Evidence for BINA as the hydrochloride form reported in the paper, not the free base/neutral form supplied as A11126; no salt equivalence, dose conversion or lot validation.
Results > Loss of functional integrity and dynamics of mGlu2 in IGEPAL (S87, S89, S90, S91); Methods > Transfection and labeling (S121); Methods > smFRET measurements (S163, S164, S165); Fig. 4 legend (S220)
VFT activation: VFT activation; comparator: Glu was sufficient to totally stabilize the receptors in the active VFT state — Results > Loss of functional integrity and dynamics of mGlu2 in IGEPAL (S89, S91)
