ライフサイエンスコーパス: BRET

1 BRET data confirmed the role of Lys-14 and Lys-15 in arr

2 BRET levels were not altered by pretreatment with seroto

3 BRET of live HEK-293 cells transfected with the subtypes

4 BRET shifts, indicating conformational change, were dete

5 BRET should be particularly useful for testing protein i

6 BRET signals comparable to those obtained from cells coe

7 BRET signals observed between AGS4-RLuc and Galpha(i1)-Y

8 BRET signals were higher for the catalytically inactive

9 BRET studies showed that, like the secretin receptor, bo

10 BRET was applied in conjunction with site-directed mutag

11 BRET was measured as an indication of receptor oligomeri

12 BRET(2) provides a better matched Forster distance to th

13 BRET(50) (BRET(50) represents the relative affinity as a

14 bioluminescence resonance energy transfer 2 (BRET(2)) in STHdh(Q7/Q7) cells.

15 ioluminescence resonance energy transfer(2) (BRET(2)) system showed a 30% increase in the BRET ratio

16 A total of 22 BRET assays have been established for nine RTKs derived

17 vior in an improved competition-based type-3 BRET assay designed to circumvent such artifacts.

18 BRET(50) (BRET(50) represents the relative affinity as acceptor/do

19 A BRET (bioluminescence resonance energy transfer) assay r

20 A BRET assay suggested that DOR phosphorylation promotes r

21 A BRET increase was observed exclusively for the gastric i

22 A BRET-based competition binding assay with 4 was also est

23 te studies of cAMP regulation we developed a BRET (bioluminescence resonance energy transfer) sensor

24 Here we present a BRET approach to monitor ligand binding to G protein-cou

25 uc and alpha2AAR-Galphai2YFP(C352I):AGS4Rluc BRET was not altered by PT treatment or Gbetagamma antag

26 FRET and BRET approaches are well established for detecting ligan

27 raTM measurements in bacterial membranes and BRET measurements made on corresponding RAGE constructs

28 orescent proteins, TagRFP and TurboFP635, as BRET acceptors.

29 QDs) are particularly well suited for use as BRET acceptors due to their high quantum yields, large S

30 fluorescent proteins are frequently used as BRET acceptors, both small molecule dyes and nanoparticl

31 e (RLuc) variants RLuc8 and RLuc8.6, used as BRET donors, combined with two red fluorescent proteins,

32 We found basal BRET interactions for some of the receptor combinations

33 m of this interaction using bioluminescence (BRET) and fluorescence (FRET) resonance energy transfer

34 mbled CB1-Galphai complexes were detected by BRET(2) Arachidonyl-2'-chloroethylamide (ACEA), a select

35 Analysis of the binding kinetics by BRET revealed remarkably long intracellular residence ti

36 ned the ability to dimerize, as monitored by BRET, whereas other mutations inhibited both nuclear exc

37 ween CheZ and CheY-P, the quantity sensed by BRET.

38 ressed with wild-type receptor, are shown by BRET(2) to heterodimerize, accounting for their dominant

39 e PRKAR1A from the catalytic PKA subunits by BRET assay.

40 refore developed a full-spectral three-color BRET assay for analyzing the specific activation of each

41 ber of beta-adrenergic agonists by comparing BRET assays of receptor-transducer interactions with Gs,

42 In contrast, BRET between tightly-associating control proteins does n

43 Conventional BRET analyses are generally done at maximal expression l

44 omers on the basis of previous, conventional BRET experiments.

45 eins versus beta-arrestin recruitment in D2R-BRET functional assays.

46 produced no ligand-independent or -dependent BRET.

47 med Arabidopsis or tobacco, we then detected BRET between three pairs of candidate interaction partne

48 DHHC3 BRET in cell membranes was decreased by the addition of

49 EGFR BRET assays are highly sensitive to known EGFR ligands [

50 We applied EGFR BRET assays to study the characteristics of somatic EGFR

51 Our results demonstrate that EGFR BRET assays precisely measure the pharmacology and signa

52 Furthermore, we demonstrate that the EGFR BRET assays are a useful tool to study the pharmacology

53 Epidermal growth factor receptor (EGFR) BRET structure-function studies identify the tyrosine re

54 cant resonance transfer signals using either BRET or a morphological FRET assay, further supporting t

55 ptors reduce but do not completely eliminate BRET transfer between receptors.

56 These red light-emitting BRET systems have great potential for investigating PPIs

57 light intensity in comparison with existing BRET fusion proteins.

58 First, BRET demonstrated a much reduced efficacy for salmeterol

59 his review examines the potential of QDs for BRET-based bioassays and imaging, and highlights example

60 Currently, FRET/BRET assays rely on co-expression of GPCR and G protein,

61 FRET signal comparable to co-expressed FRET/BRET sensors.

62 Functional BRET assays included activation of G proteins with all G

63 Thus, ER homodimer and heterodimer BRET assays are applicable to drug screening for dimer-s

64 4 enabled the establishment of a homogeneous BRET-based binding assay suitable for both detailed kine

65 uses enzyme-catalyzed luminescence; however, BRET signals usually have been too dim to image effectiv

66 fic interaction as indicated by a hyperbolic BRET signal in response to increasing PAR4-GFP expressio

67 The inhibitory regulation of GPR-Galpha(i1) BRET by Ric-8A was blocked by pertussis toxin.

68 The enhancement of GPR-Galpha(i1) BRET observed with Ric-8A was further augmented by pertu

69 In BRET experiments, the PAR4 homodimers showed a specific

70 er domains, resulting in a large decrease in BRET efficiency.

71 Such key improvement in BRET measurement paves the way for the simultaneous moni

72 st, caused a rapid and transient increase in BRET efficiency (BRETEff) between Galphai-Rluc and CB1-g

73 astrin-4) resulted in the rapid reduction in BRET signal in contrast to the enhancement of such a sig

74 Agonist-induced reduction in BRET signal was also observed for pairs of CCK receptors

75 ation, and this agonist-induced reduction in BRET was blocked by pertussis toxin.

76 PAR4 did not interact with rhodopsin in BRET assays.

77 Deletion of S2 produced ligand-independent BRET for only those pairings normally occurring in the p

78 Ligand-induced BRET changes assessing Gbetagamma-Kir3.1 subunit interac

79 at the standard frameworks used to interpret BRET titration experiments rely on simplifying assumptio

80 Here, using an intramolecular BRET (bioluminescence resonance energy transfer)-based b

81 ophysical experiments with an intramolecular BRET beta-arrestin2 biosensor revealed that osmotic stre

82 Even more intriguing, BRET experiments indicate that CTAP (D-Phe-Cys-Tyr-D-Trp

83 e pharmacological characterization involving BRET biosensors, binding studies, electrophysiology, and

84 A lower BRET(50) for the alpha(2A)-alpha(2C) heterodimer (0.79 +

85 r/donor ratio required to reach half-maximal BRET [bioluminescence resonance energy transfer] values)

86 The threshold maximum BRET signal was disrupted in a concentration-dependent m

87 R-Kir3 interactions unmodified but modulated BRET between DOR-GalphaoA, DOR-Gbetagamma, GalphaoA-Gbet

88 h the K2A mutations had little effect on net BRET(max) values for the M2 muscarinic acetylcholine (M2

89 Future development of new BRET acceptors should further expand the multiplexing ca

90 The imaging utility of the new BRET vector is shown by constructing a sensor using two

91 This new BRET fusion protein (BRET3) exhibits severalfold improve

92 This new BRET strategy provides a unique platform to assay protei

93 This new BRET vector should facilitate high-throughput sensitive

94 This new BRET vector shows an overall 5.5-fold improvement in the

95 s, as well as a high specific to nonspecific BRET binding signal.

96 efficiency, we report generation of a novel BRET vector by fusing a GFP(2) acceptor protein with a n

97 ing did not significantly alter the observed BRET(2) signal, suggesting that CXCR4 exists as a consti

98 The advantages of BRET include expressing full-length proteins in their na

99 r results demonstrate a novel application of BRET for assessing target engagement within the complex

100 The CCD imaging approach of BRET signal is particularly appealing due to its capacit

101 urther expand the multiplexing capability of BRET and improve its applicability and sensitivity for i

102 The combination of BRET/bimolecular luminescence complementation assay reve

103 The dependence of BRET efficiency on acceptor/donor ratio at fixed surface

104 multaneous visualization and quantitation of BRET signal from live cells and cells implanted in livin

105 ng advantage of the critical relationship of BRET efficiency and donor quantum efficiency, we report

106 associated with the nonrigorous treatment of BRET data are illustrated for the case of G protein-coup

107 multimerization as well as a new variant of BRET assay that is useful for measuring the interactions

108 Our BRET study also confirmed that: (1) capsaicin and heat p

109 lly-induced recruitment of local third-party BRET donors or acceptors reliably separates nonspecific

110 VPAC2, and secretin receptors, and performed BRET and morphologic fluorescence resonance energy trans

111 tion was induced, as indicated by a positive BRET signal, on exposure of the cells to bivalent ligand

112 tion-mediated desensitization, also produced BRET signals above background.

113 s and imaging, and highlights examples of QD-BRET for biosensing and imaging applications.

114 ioluminescence resonance energy transfer (QD-BRET) to detect the protease activity in complex biologi

115 However, quantitative BRET(2) analyses in intact cells indicate a lack of effe

116 inated the disruptive effect on CCK receptor BRET, whereas the other mutant peptide behaved like wild

117 that receptor had no effect on CCK receptor BRET.

118 ly TM VI and VII peptides disrupted receptor BRET.

119 ormation within complexes, reducing receptor BRET signals.

120 drenergic receptor (beta2AR), and reevaluate BRET titration as a method to study membrane protein ass

121 and active H-Ras(G/V)-Venus exhibit a robust BRET signal at the plasma membrane that is markedly enha

122 4-Renilla luciferase (Rluc) exhibited robust BRET with the tethered GalphaiYFP, and this interaction

123 RTK BRET assays are highly sensitive for quantifying ligand-

124 RTK BRET-2 assays monitor, in living cells, the specific int

125 -terminus of GLUT1 and performing saturation BRET analysis, we were able to demonstrate the formation

126 should facilitate high-throughput sensitive BRET assays, including studies in single live cells and

127 the first time that an efficient sequential BRET-FRET energy transfer process based on firefly lucif

128 n the constitutive presence of a significant BRET signal above that in a series of controls, with thi

129 hough all constructs generated a significant BRET signal, this was disrupted by peptide in all except

130 A similar BRET(2)-based transduction scheme approach would likely

131 In addition, specific BRET signals were observed for AGS4-RLuc and alpha(2)-ad

132 reliably separates nonspecific and specific BRET.

133 otein (YFP) demonstrated saturable, specific BRET signals.

134 sence and absence of rapamycin, the specific BRET signal was determined.

135 ed to an image splitter, we demonstrate that BRET can be used to image protein interactions in plant

136 Further, the fact that BRET changes at the Gbetagamma-Kir3 interface are predic

137 iple levels of donor expression we find that BRET between beta2AR protomers is directly proportional

138 The BRET assay, based on the interaction with Calmodulin, wa

139 The BRET technique is complementary to one based on FRET, de

140 The BRET technology provides an assay platform to study sign

141 Additionally, the BRET assay suggested that depletion of phosphorylation d

142 teins as the BRET donor, quantum dots as the BRET acceptor, and protease substrates sandwiched betwee

143 rs consist of bioluminescent proteins as the BRET donor, quantum dots as the BRET acceptor, and prote

144 We concluded the BRET assay is an accurate, sensitive, and cost/time effi

145 ontrast, rhodopsin was unable to disrupt the BRET signal, indicating that the disruption of the PAR4

146 cells with brefeldin A did not eliminate the BRET signals, and morphologic FRET experiments confirmed

147 anging from 10nM to 3.16 muM maltose for the BRET(2) system compared to an EC(50) of 2.3 muM and a li

148 However, the BRET vectors currently used lack adequate sensitivity fo

149 her, chemicals that screened positive in the BRET assay also stimulated phenotypic outcomes in daphni

150 BRET(2)) system showed a 30% increase in the BRET ratio upon maltose binding, compared with a 10% inc

151 shows an overall 5.5-fold improvement in the BRET ratio, thereby greatly enhancing the dynamic range

152 troduction of a single point mutation in the BRET(2) tagged MBP protein.

153 sites on surface enabled us to maximize the BRET efficiency by adjusting the QD/enzyme conjugation r

154 the absence of ligand, we have monitored the BRET signal after deletion of regions of the extracellul

155 The specificity of the BRET and FRET signals was confirmed by control studies.

156 nical advances to enhance the utility of the BRET assay in plants.

157 y greatly enhancing the dynamic range of the BRET signal.

158 In addition, the low background of the BRET system allowed detection of significant, but less e

159 similarly observed to have no effect on the BRET(2) signal.

160 Overall, the BRET assay had comparable or greater sensitivity as comp

161 ) clock genes from cyanobacteria, we use the BRET technique to demonstrate that the clock protein Kai

162 Using the BRET-based biosensor, maltose in water was detected on a

163 rhodopsin are quantified and imaged with the BRET Ca(++) sensor in darkness, thereby avoiding undesir

164 gands in mice showed no correlation with the BRET data is consistent with the absence of association

165 These BRET systems consist of the recently developed Renilla r

166 We show the use of these BRET systems for ratiometric imaging of both cells in cu

167 , bioluminescence resonance energy transfer (BRET(2)) analyses confirmed that the hLHR constitutively

168 g bioluminescence resonance energy transfer (BRET(2)), we demonstrated that CXCR4 multimers are found

169 a bioluminescence resonance energy transfer (BRET) and enabled the complex to emit NIR light.

170 a bioluminescence resonance energy transfer (BRET) approach, we demonstrate that a series of antipsyc

171 d bioluminescence resonance energy transfer (BRET) assay platform, our studies in human embryonic kid

172 minescent Forster resonance energy transfer (BRET) assay using the luminescent donor Nanoluciferase a

173 e bioluminescence resonance energy transfer (BRET) assays for monitoring the formation of ERalpha/bet

174 nd bioluminescent resonance energy transfer (BRET) assays in transfected cells, the present study att

175 y bioluminescence resonance energy transfer (BRET) assays of live cells.

176 d bioluminescence resonance energy transfer (BRET) assays of membrane protein stoichiometry, the pres

177 , bioluminescence resonance energy transfer (BRET) assays revealed that sustained activation by SNC-8

178 d bioluminescence resonance energy transfer (BRET) assays were developed to monitor the activation of

179 e bioluminescence resonance energy transfer (BRET) assays.

180 y bioluminescence resonance energy transfer (BRET) assays.

181 n bioluminescence resonance energy transfer (BRET) between JNK3-luciferase and Venus-arrestins.

182 f bioluminescence resonance energy transfer (BRET) between receptor constructs that included carboxyl

183 Bioluminescence resonance energy transfer (BRET) between Renilla luciferase and yellow fluorescent

184 y bioluminescence resonance energy transfer (BRET) between Renilla luciferase fused to the phosphatas

185 r bioluminescence resonance energy transfer (BRET) between RLuc8 and iRFPs, the chimeric luciferases

186 ed bioluminescent resonance energy transfer (BRET) biosensor, comprising maltose binding protein (MBP

187 , bioluminescence resonance energy transfer (BRET) biosensors, and the label-free approach surface pl

188 h bioluminescence resonance energy transfer (BRET) donor/acceptor pairs that allowed us to evaluate i

189 Bioluminescence resonance energy transfer (BRET) experiments revealed that DHHC2 or DHHC3 (Golgi-sp

190 y bioluminescence resonance energy transfer (BRET) in human embryonic kidney 293 cells.

191 d bioluminescence resonance energy transfer (BRET) in live cells to identify a short motif in the C-t

192 g bioluminescence resonance energy transfer (BRET) in live cells, we show that RGS14-Luciferase and a

193 g bioluminescence resonance energy transfer (BRET) in live cells, we show that WNT5A stimulates dimer

194 Bioluminescence resonance energy transfer (BRET) is a natural biophysical phenomenon that underlies

195 Bioluminescence resonance energy transfer (BRET) is a well-established method for investigating pro

196 Bioluminescence resonance energy transfer (BRET) is currently used for monitoring various intracell

197 Bioluminescence resonance energy transfer (BRET) is often used to study association of membrane pro

198 t bioluminescence resonance energy transfer (BRET) occurred minimally in intact versions of these rec

199 Bioluminescence resonance energy transfer (BRET) operates with biochemical energy generated by biol

200 e bioluminescence resonance energy transfer (BRET) phenomenon, we report the development of a highly

201 A bioluminescence resonance energy transfer (BRET) readout of heterotrimer activation with high tempo

202 ) bioluminescence resonance energy transfer (BRET) reporters to monitor conformational changes in bet

203 h bioluminescence resonance energy transfer (BRET) studies of liganded-beta2AR binding to arrestin an

204 d bioluminescence resonance energy transfer (BRET) studies on COS cells coexpressing MOP and CCK2 rec

205 d bioluminescence resonance energy transfer (BRET) studies.

206 a bioluminescence resonance energy transfer (BRET) system, and split reporter protein complementation

207 d bioluminescence resonance energy transfer (BRET) techniques, calcium flux measurements, and microsc

208 e bioluminescence resonance energy transfer (BRET) technology to quantitatively study the pharmacolog

209 e bioluminescence resonance energy transfer (BRET) technology, which directly measures the recruitmen

210 n bioluminescence resonance energy transfer (BRET) that allow for assaying PPIs both in cell culture

211 n bioluminescence resonance energy transfer (BRET) that allows detection of antibodies directly in so

212 d bioluminescence resonance energy transfer (BRET) to demonstrate that the prototypic family B secret

213 d bioluminescence resonance energy transfer (BRET) to detect and quantify assembly of the methyl farn

214 d bioluminescence resonance energy transfer (BRET) to examine oligomerization of Ste2p, a G protein-c

215 g bioluminescence resonance energy transfer (BRET) to reveal the binding characteristics of a drug wi

216 e bioluminescence resonance energy transfer (BRET) to show that after activation, Galphas rapidly ass

217 d bioluminescence resonance energy transfer (BRET) to study the Arf1/AP-1 interaction and AP-1 confor

218 Bioluminescence resonance energy transfer (BRET) was assessed in HEK293 cells expressing 5-HT(2C) r

219 d bioluminescence resonance energy transfer (BRET) were used to examine the PAR4 homodimer interface.

220 , bioluminescence resonance energy transfer (BRET), and functional analysis to map spatial approximat

221 , bioluminescence resonance energy transfer (BRET), avoids these problems because it uses enzyme-cata

222 d bioluminescence resonance energy transfer (BRET), uses a bioluminescent luciferase that is genetica

223 g bioluminescence resonance energy transfer (BRET), we detected a constitutive and phorbol 12-myrista

224 Bioluminescence resonance energy transfer (BRET), which relies on nonradiative energy transfer betw

225 a bioluminescence resonance energy transfer (BRET)-based assay.

226 n bioluminescence resonance energy transfer (BRET)-based assays to give an insight into the structure

227 r bioluminescence resonance energy transfer (BRET)-based biosensor, capable of detecting signal-depen

228 y bioluminescence resonance energy transfer (BRET)-based saturation and kinetic binding experiments,

229 l bioluminescence resonance energy transfer (BRET)-fluorescence resonance energy transfer (FRET) proc

230 g bioluminescence resonance energy transfer (BRET).

231 g bioluminescence resonance energy transfer (BRET).

232 f bioluminescence resonance energy transfer (BRET).

233 y bioluminescence resonance energy transfer (BRET).

234 d bioluminescence resonance energy transfer (BRET-2) assays, diltiazem was a partial agonist at GHSR1

235 We have used BRET to study the in vivo activation of AP-1.

236 al approach to test this hypothesis, we used BRET to examine 7TM receptor-mediated regulation of Galp

237 Using BRET-based biosensors, we showed that whereas K17F activ

238 -protein interaction can be documented using BRET.

239 Finally we provide conclusive evidence using BRET and FRET that OXRs and GPR103 form functional heter

240 ions between CB1 and D2L were observed using BRET(2) Cotreatment of STHdh(Q7/Q7) cells with ACEA and

241 This work validates BRET as a powerful tool for interrogating and observing

242 beta-arrestin recruitment (as monitored via BRET assays).

243 assessing membrane protein association with BRET.

244 tor and GIP receptor were characterized with BRET donor saturation studies, shift experiments, and te

245 alpha(i1)-YFP and AGS4-Rluc-G-Galpha(i1)-YFP BRET were observed in both pellet and supernatant subcel

246 -Galpha(i1)-YFP and AGS4-Rluc-Galpha(i1)-YFP BRET were regulated by Ric-8A but not by Galpha-interact