2013
DOI: 10.1371/journal.pone.0062247
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A High Throughput Screen for RGS Proteins Using Steady State Monitoring of Free Phosphate Formation

Abstract: G-protein coupled receptors are a diverse group that are the target of over 50% of marketed drugs. Activation of these receptors results in the exchange of bound GDP for GTP in the Gα subunit of the heterotrimeric G-protein. The Gα subunit dissociates from the β/γ subunits and both proceed to affect downstream signaling targets. The signal terminates by the hydrolysis of GTP to GDP and is temporally regulated by Regulators of G-protein Signaling (RGS) proteins that act as GTPase Activating Proteins (GAPs). Thi… Show more

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Cited by 31 publications
(62 citation statements)
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“…The ultimate colorimetric complex is formed after the formation of phosphomolybdate, indicating enough time is required for full color development [23]. Hence, the staining time was optimized (Fig.…”
Section: Optimization Of the Parameters Of Malachite Green Methodsmentioning
confidence: 99%
“…The ultimate colorimetric complex is formed after the formation of phosphomolybdate, indicating enough time is required for full color development [23]. Hence, the staining time was optimized (Fig.…”
Section: Optimization Of the Parameters Of Malachite Green Methodsmentioning
confidence: 99%
“…RGS17 is ubiquitously expressed, from yeast to mammals but it is expressed in few tissues and organs (Bodle et al, 2013;Jung et al, 2012;Mackie and Roman, 2011;Monroy et al, 2013b;Mosakhani et al, 2013;Zhang et al, 2012a). It is expressed mainly in brain areas heavily innervated with dopamine, such as the nucleus accumbens and striatum (Stanwood et al, 2006) and high significance in the cerebellum (Larminie et al, 2004) and in the central nervous system, in the hypothalamus, midbrain, and pons-medulla (Garzon et al, 2005a).…”
Section: Rgs17 (Rgsz2)mentioning
confidence: 99%
“…It is generally known that GDP/GTP exchange is the rate-limiting step in multiple turnover measurements of G␣ isoforms (9 -12). Therefore, beside steady-state assays using ␥-32 P labeling (13) or malachite green (14), pre-steady-state assays are used to characterize the hydrolysis reaction of G␣ isoforms (15)(16)(17)(18). We present here for the first time single turnover measurements of G␣ i1 using time-resolved FTIR spectroscopy, an ultrasensitive method that can be applied in solution and has been successfully used for photoactivable proteins like bacteriorhodopsin (19,20), channelrhodopsin (21), and other rhodopsins (22).…”
mentioning
confidence: 99%