Experimental Determination of the Förster Distance for Two Commonly Used Bioluminescent Resonance Energy Transfer Pairs

Dacres, Helen; Wang, J.; Dumancic, Mira M.; Trowell, Stephen

doi:10.1021/ac9022956

Cited by 55 publications

(49 citation statements)

References 24 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The use of BRET pairs may also magnify this effect. The r 0 for the BRET pair used in BRET 2 experiments is 7.5 nm; the largest value reported for a resonance energy transfer pair used in biological studies (67); and, as shown in Fig. 6, the effect of protein density on FRET efficiency is greater for energy transfer pairs with larger r 0 .…”

Section: Discussionmentioning

confidence: 86%

Restricted Lateral Diffusion of Luteinizing Hormone Receptors in Membrane Microdomains

Wolf‐Ringwall

Winter

Liu

et al. 2011

Journal of Biological Chemistry

View full text Add to dashboard Cite

Signal transduction by luteinizing hormone (LH)3 receptor plays an essential role in the normal reproductive function of both male and female mammals by promoting ovulation, follicle maturation, corpus luteum formation, and steroidogenesis. LH receptor signaling in response to saturating concentrations of LH or human chorionic gonadotropin (hCG) involves substantial changes in receptor lateral and rotational dynamics (1, 2) as well as the association of LH receptors with membrane rafts (3). FRET techniques (4) and electron microscopy (5) indicate that functional hormone-receptor complexes can also become self-associated into dimers/oligomers following ligand binding. Previous experimental strategies have examined changes in receptor motions on collections of cells or large numbers of fluorescently tagged molecules on single cells. It is now possible, however, to examine the lateral motions of individual LH receptors on living cells using microscope-based single particle tracking techniques (6).Although the mechanism involved in retention of hCG-occupied LH receptors in small membrane compartments is still unclear, exposure to saturating concentrations of hCG results in confinement of the majority of LH receptors within small cell surface compartments. LH receptors remain within these compartments for comparatively long times and appear to diffuse pseudo-randomly before being captured within another compartment of similar size (7). Similar behavior has been described and analyzed by Kusumi and co-workers (6) for selected phospholipids and for transferrin receptors (8) and by Daumas and co-workers (9) for the -opioid receptor. Daumas argues that the -opioid receptor can both diffuse within the bulk membrane and exhibit confinement within a microdomain that itself diffuses slowly. Kusumi and co-workers (6) suggest that particles may be confined by proteins forming a barrier that is either continuous or discontinuous, leading to receptor diffusion within small membrane regions accompanied by intermittent escape from compartments and periods of unrestricted diffusion in the bulk membrane.Our previous studies of LH receptor lateral and rotational diffusion suggest that actin microfilaments or membrane protein interactions with the cytoskeleton may provide organizing structures that restrict the lateral motions of receptors (1, 2). However, the mechanism of hormone-initiated LH receptor confinement in small compartments has not been examined previously in detail at the single molecule level. In the present work, we compared the lateral dynamics of individual LH receptors that were either FLAG-tagged and identified using FLAG-specific antibodies with the diffusion of native receptors occupied by gold nanoparticle-hCG conjugates (Au-hCG) or gold-deglycosylated hCG (Au-DG-hCG). The latter material is

show abstract

Section: Discussionmentioning

confidence: 86%

Restricted Lateral Diffusion of Luteinizing Hormone Receptors in Membrane Microdomains

Wolf‐Ringwall

Winter

Liu

et al. 2011

Journal of Biological Chemistry

View full text Add to dashboard Cite

show abstract

“…The efficiency of energy transfer depends on the relative orientation of the donor and acceptor and the distance between them (Zacharias et al, 2000), so that absolute distances can not be measured. Experimentally determined Förster distance R 0 (distance at which the energy transfer efficiency is 50%) for BRET 1 and BRET 2 is 4.4 nm and 7.5 nm, respectively (Dacres et al, 2010). 7TMR transmembrane core spans ~40 Å across the intracellular surface (Palczewski et al, 2000), which makes BRET suitable to the study of dimerization.…”

Section: Interpretation Of Bret Results -Possible Drawbacksmentioning

confidence: 99%

“…where r is a distance between donor and acceptor, Förster radius R 0 depends on spectral overlap and dipole orientations yielding R 0 values of 4.4 nm for BRET 1 and 7.5 nm for BRET 2 (Dacres et al, 2010). E is an important parameter in interpretation of the BRET assays used for oligomerisation studies.…”

Section: Basic Assumptionsmentioning

confidence: 99%

Quantitative Assessment of Seven Transmembrane Receptors (7TMRs) Oligomerization by Bioluminescence Resonance Energy Transfer (BRET) Technology

Kubale¹,

Drinovec²,

Vrecl³

2012

Bioluminescence - Recent Advances in Oceanic Measurements and Laboratory Applications

View full text Add to dashboard Cite

“…The BRET 2 system is reported to produce much less light than the BRET H system due to the fact that quantum yield of coelenterazine 400a is two order of magnitude smaller than that of coelenterazine h. 18 However, because of its large Förster distance and spectral separation, 11 BRET 2 is ideally matched to the size of the thrombin biosensor. Accordingly, the BRET 2 -based thrombin sensor is a good option for validating the microfluidic setup and optical detection system for high sensitivity biosensing.…”

Section: Experimental a Materialsmentioning

confidence: 99%

“…[5][6][7][8][9][10] BRET 2 is ideally matched to the scale of many protein interactions and rearrangements due to its relatively long Förster distance but has relatively low luminance arising from the low quantum yield of the substrate oxidation. 11 Recently, we reported a microscope-based microfluidic system for BRET-based biosensing, 12 , we made two changes to the design of the detection system. First, we increased the volume of reaction that is optically sampled by incorporating a cylindrical chamber ("BRET reaction chamber"), perpendicular to the common direction of flow with a total volume of 2p ll.…”

Section: Introductionmentioning

confidence: 99%

Sub-nanomolar detection of thrombin activity on a microfluidic chip

Gel

Dacres

et al. 2014

Biomicrofluidics

View full text Add to dashboard Cite

Bioluminescence resonance energy transfer (BRET) is a form of Förster resonance energy transfer. BRET has been shown to support lower limits of detection than fluorescence resonance energy transfer (FRET) but, unlike FRET, has not been widely implemented on microfluidic devices for bioanalytical sensing. We recently reported a microscope-based microfluidic system for BRET-based biosensing, using a hybrid, high quantum-efficiency, form of BRET chemistry. This paper reports the first optical fiber-based system for BRET detection on a microfluidic chip, capable of quantifying photon emissions from the low quantum-efficiency BRET 2 system. We investigated the effects of varying core diameter and numerical aperture of optical fibers, as well as varying microfluidic channel design and measurement conditions. We optimized the set-up in order to maximize photon counts and minimize the response time. The optimized conditions supported measurement of thrombin activity, with a limit of detection of 20 pM, which is lower than the microscope-based system and more than 20 times lower than concentrations reported to occur in plasma clots. V C 2014 AIP Publishing LLC.

show abstract

Experimental Determination of the Förster Distance for Two Commonly Used Bioluminescent Resonance Energy Transfer Pairs

Cited by 55 publications

References 24 publications

Restricted Lateral Diffusion of Luteinizing Hormone Receptors in Membrane Microdomains

Restricted Lateral Diffusion of Luteinizing Hormone Receptors in Membrane Microdomains

Quantitative Assessment of Seven Transmembrane Receptors (7TMRs) Oligomerization by Bioluminescence Resonance Energy Transfer (BRET) Technology

Sub-nanomolar detection of thrombin activity on a microfluidic chip

Contact Info

Product

Resources

About