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BioAssay: AID 602148

Active GTPase Screen Compounds affects on binding of VLA-4 specific ligand

Activation of GTPases (in particular Rho family) are involved in the signal transduction cascades that commences with stimulation of formyl peptide receptors and resulting in the increase ligand-affinity of VLA-4 integrins. Assessment of this increased affinity can be made with , fluorescently labeled specific ligand for VLA-4, LDV-FITC. By utilizing U937 cells expressing non-desensetizing, more ..
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 Tested Compounds
 Tested Compounds
All(20)
 
 
Active(10)
 
 
Inactive(10)
 
 
 Tested Substances
 Tested Substances
All(20)
 
 
Active(10)
 
 
Inactive(10)
 
 
AID: 602148
Data Source: NMMLSC (VLA4_LDVFITC_U937_Cells_GTPaseCompounds )
Depositor Category: NIH Molecular Libraries Probe Production Network
Deposit Date: 2011-12-12
Hold-until Date: 2012-12-05
Modify Date: 2012-12-05

Data Table ( Complete ):           View Active Data    View All Data
Target
BioActive Compounds: 10
Related Experiments
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AIDNameTypeProbeComment
1772Project utilizing multiplex HTS to identify specific small molecule inhibitors of Ras and Ras-related GTPasesSummary5 depositor-specified cross reference: Summary report for GTPase target project
757HTS to identify specific small molecule inhibitors of Ras and Ras-related GTPases specifically Rac wildtypeScreening same project related to Summary assay
758HTS to identify specific small molecule inhibitors of Ras and Ras-related GTPases specifically Rab7 wildtypeScreening same project related to Summary assay
759HTS to identify specific small molecule inhibitors of Ras and Ras-related GTPases specifically Ras wildtypeScreening same project related to Summary assay
760HTS to identify specific small molecule inhibitors of Ras and Ras-related GTPases specifically Rab2 wildtypeScreening same project related to Summary assay
761HTS to identify specific small molecule inhibitors of Ras and Ras-related GTPases specifically Cdc42 wildtypeScreening same project related to Summary assay
764HTS to identify specific small molecule inhibitors of Ras and Ras-related GTPases specifically Rac activated mutantScreening same project related to Summary assay
1333Multiplex dose response to identify specific small molecule inhibitors of Ras and Ras-related GTPases specifically Cdc42 activated mutantConfirmatory same project related to Summary assay
1334Multiplex dose response to identify specific small molecule inhibitors of Ras and Ras-related GTPases specifically Cdc42 wildtypeConfirmatory same project related to Summary assay
1335Multiplexed dose response to identify specific small molecule inhibitors of Ras and Ras-related GTPases specifically Ras wildtypeConfirmatory same project related to Summary assay
1336Multiplex dose response to identify specific small molecule inhibitors of Ras and Ras-related GTPases specifically Rab7 wildtypeConfirmatory same project related to Summary assay
1337Multiplex dose response to identify specific small molecule inhibitors of Ras and Ras-related GTPases specifically Rab2 wildtypeConfirmatory same project related to Summary assay
1339Multiplex dose response to identify specific small molecule inhibitors of Ras and Ras-related GTPases specifically Rac activated mutantConfirmatory same project related to Summary assay
1340Multiplexed dose response to identify specific small molecule inhibitors of Ras and Ras-related GTPases specifically Rac wildtypeConfirmatory same project related to Summary assay
1341Multiplexed dose response to identify specific small molecule inhibitors of Ras and Ras-related GTPases specifically Ras activated mutantConfirmatory same project related to Summary assay
1758Activator binding kinetics on Ras and Ras-related GTPases, specifically Cdc42_wtScreening same project related to Summary assay
1759Activator binding kinetics on Ras and Ras-related GTPases, specifically Ras_wtScreening same project related to Summary assay
1760Activator binding kinetics on Ras and Ras-related GTPases, specifically Rab7_wtScreening same project related to Summary assay
1761Activator binding kinetics on Ras and Ras-related GTPases, specifically Ras_act (activated mutant)Screening same project related to Summary assay
1762Activator binding kinetics on Ras and Ras-related GTPases, specifically Cdc42_act (activated mutant)Screening same project related to Summary assay
1763Activator binding kinetics on Ras and Ras-related GTPases, specifically Rab2_wtScreening same project related to Summary assay
1769Profiling Assay to determine GST-GSH interactions in multiplex bead-based assaysConfirmatory same project related to Summary assay
2009Oxadiazole SAR compounds tested by Multiplex dose response to identify specific small molecule inhibitors of Ras and Ras-related GTPases specifically Cdc42 activated mutantConfirmatory same project related to Summary assay
2019Pyrazoline SAR compounds tested via Multiplex dose response to identify specific small molecule inhibitors of Ras and Ras-related GTPases specifically Cdc42 wildtype proteinConfirmatory same project related to Summary assay
2020Pyrazoline SAR compounds tested via Multiplex dose response to identify specific small molecule inhibitors of Ras and Ras-related GTPases specifically Cdc42 activated mutantConfirmatory same project related to Summary assay
2021Additional SAR compounds tested by Multiplex dose response to identify specific small molecule inhibitors of Ras and Ras-related GTPases specifically Cdc42 activated mutantConfirmatory same project related to Summary assay
2022Additional SAR compounds tested via Multiplex dose response to identify specific small molecule inhibitors of Ras and Ras-related GTPases specifically Cdc42 wildtypeConfirmatory same project related to Summary assay
2027Oxadiazole SAR compounds tested via Multiplex dose response to identify specific small molecule inhibitors of Ras and Ras-related GTPases specifically Rac1 wildtypeConfirmatory same project related to Summary assay
2031Oxadiazole SAR compounds tested via Multiplex dose response to identify specific small molecule inhibitors of Ras and Ras-related GTPases specifically Rab7 wildtypeConfirmatory same project related to Summary assay
2033Oxadiazole SAR compounds tested via Multiplex dose response to identify specific small molecule inhibitors of Ras and Ras-related GTPases specifically Rab2 wildtypeConfirmatory same project related to Summary assay
2036Additional SAR compounds tested via Multiplex dose response to identify specific small molecule inhibitors of Ras and Ras-related GTPases specifically Rab7 wildtypeConfirmatory same project related to Summary assay
2037Oxadiazole SAR compounds tested via Multiplex dose response to identify specific small molecule inhibitors of Ras and Ras-related GTPases specifically Cdc42 wildtypeConfirmatory same project related to Summary assay
2038Additional SAR compounds tested via Multiplex dose response to identify specific small molecule inhibitors of Ras and Ras-related GTPases specifically Ras wildtypeConfirmatory same project related to Summary assay
2039Additional SAR compounds tested via Multiplex dose response to identify specific small molecule inhibitors of Ras and Ras-related GTPases specifically Rac1 activated mutantConfirmatory same project related to Summary assay
2040Additional SAR compounds tested via Multiplex dose response to identify specific small molecule inhibitors of Ras and Ras-related GTPases specifically Rac1 wildtypeConfirmatory same project related to Summary assay
2041Pyrazoline SAR compounds tested via Multiplex dose response to identify specific small molecule inhibitors of Ras and Ras-related GTPases specifically Rab7 wildtypeConfirmatory same project related to Summary assay
2042Oxadiazole SAR compounds tested via Multiplex dose response to identify specific small molecule inhibitors of Ras and Ras-related GTPases specifically Ras activated mutantConfirmatory same project related to Summary assay
2043Additional SAR compounds tested via Multiplex dose response to identify specific small molecule inhibitors of Ras and Ras-related GTPases specifically Ras activated mutantConfirmatory same project related to Summary assay
2045Pyrazoline SAR compounds tested via Multiplex dose response to identify specific small molecule inhibitors of Ras and Ras-related GTPases specifically Rab2 wildtypeConfirmatory same project related to Summary assay
2046Additional SAR compounds tested via Multiplex dose response to identify specific small molecule inhibitors of Ras and Ras-related GTPases specifically Rab2 wildtypeConfirmatory same project related to Summary assay
2047Pyrazoline SAR compounds tested via Multiplex dose response to identify specific small molecule inhibitors of Ras and Ras-related GTPases specifically Ras wildtypeConfirmatory same project related to Summary assay
2048Pyrazoline SAR compounds tested via Multiplex dose response to identify specific small molecule inhibitors of Ras and Ras-related GTPases specifically Rac1 activated mutantConfirmatory same project related to Summary assay
2050Pyrazoline SAR compounds tested via Multiplex dose response to identify specific small molecule inhibitors of Ras and Ras-related GTPases specifically Ras activated mutantConfirmatory same project related to Summary assay
2051Oxadiazole SAR compounds tested via Multiplex dose response to identify specific small molecule inhibitors of Ras and Ras-related GTPases specifically Rac1 activated mutantConfirmatory same project related to Summary assay
2053Oxadiazole SAR compounds tested via Multiplex dose response to identify specific small molecule inhibitors of Ras and Ras-related GTPases specifically Ras wildtypeConfirmatory same project related to Summary assay
2055Pyrazoline SAR compounds tested via Multiplex dose response to identify specific small molecule inhibitors of Ras and Ras-related GTPases specifically Rac1 wildtypeConfirmatory same project related to Summary assay
2372Compound effect on equilibrium binding with Cdc42 under varying GTP conditions: nucleotide depletion with Mg bufferOther same project related to Summary assay
2373Compound effect on equilibrium binding with Cdc42 under varying GTP conditions with Mg bufferOther same project related to Summary assay
2374PAK-Effector Pull-down Assay for Quantification of Rac/Cdc42 Activation Using 3T3 CellsOther same project related to Summary assay
2375Panel results of Cell based ELISA Assessing Activation of Cdc42 and Rac1Other same project related to Summary assay
2376Dose Response of compounds with constant GTP under the condition of nascent nucleotide depletion and Mg bufferConfirmatory same project related to Summary assay
2378Compound effect on equilibrium binding with Cdc42 under varying GTP conditions with EDTA bufferOther same project related to Summary assay
2393Dose Response of compounds for six proteins with constant GTP under the condition of Mg bufferOther same project related to Summary assay
2418Assessment of Cdc42 inhibitors on Bradykinin activation of 3T3 cellsOther same project related to Summary assay
588369Cellular toxicity assessed by Trypan Blue for compounds active in GTPase screenOther same project related to Summary assay
588373SAR compounds for Cdc42 probe extension project tested by Multiplex dose response to identify specific small molecule inhibitors of Ras and Ras-related GTPases specifically Cdc42 activated mutant, round 1Confirmatory same project related to Summary assay
588377SAR compounds for Cdc42 probe extension project tested by Multiplex dose response to identify specific small molecule inhibitors of Ras and Ras-related GTPases specifically Cdc42 wildtype, round 1Confirmatory same project related to Summary assay
588381SAR compounds for Cdc42 probe extension project tested by Multiplex dose response to identify specific small molecule inhibitors of Ras and Ras-related GTPases specifically H-Ras activated mutant, round 1Confirmatory same project related to Summary assay
588383SAR compounds for Cdc42 probe extension project tested by Multiplex dose response to identify specific small molecule inhibitors of Ras and Ras-related GTPases specifically Ras wildtype, round 1Confirmatory same project related to Summary assay
588384SAR compounds for Rab7 project tested by Multiplex dose response to identify specific small molecule inhibitors of Ras and Ras-related GTPases specifically Cdc42 activated mutant, round 1Confirmatory same project related to Summary assay
588385SAR compounds tested by Multiplex dose response to identify specific small molecule inhibitors of Ras and Ras-related GTPases specifically Cdc42 wildtype, round 1Confirmatory same project related to Summary assay
588387SAR compounds tested by Multiplex dose response to identify specific small molecule inhibitors of Ras and Ras-related GTPases specifically H-Ras activated mutant, round 1Confirmatory same project related to Summary assay
588388SAR compounds tested by Multiplex dose response to identify specific small molecule inhibitors of Ras and Ras-related GTPases specifically H-Ras wildtype, round 1Confirmatory same project related to Summary assay
588393SAR compounds for Cdc42 probe extension project tested by Multiplex dose response to identify specific small molecule inhibitors of Ras and Ras-related GTPases specifically Rab7 wildtype, round 1Confirmatory same project related to Summary assay
588394SAR compounds tested by Multiplex dose response to identify specific small molecule inhibitors of Ras and Ras-related GTPases specifically Rab7 wildtype, round 1Confirmatory same project related to Summary assay
588410Cellular toxicity assessed by Trypan Blue for compounds in active GTPase screen, set2Other same project related to Summary assay
588427Cellular toxicity assessed by Trypan Blue for compounds in active GTPase screen, Set1Other same project related to Summary assay
588477Dose Response of round 1 SAR compounds for Cdc42 probe extension project tested by multiplex of eight GTPase proteins under the condition of Mg Buffer, non-chelator bufferOther same project related to Summary assay
588479Dose Response of round 1 SAR compounds for GTPase inhibitor project tested by multiplex of eight GTPase proteins under the condition of Mg Buffer, non-chelator bufferOther same project related to Summary assay
588622Dose Response of round 2 SAR compounds for GTPase inhibitor project tested by multiplex of eight GTPase proteins under the condition of Mg Buffer, non-chelator bufferOther same project related to Summary assay
588624SAR compounds for Rab7 project tested by Multiplex dose response to identify specific small molecule inhibitors of Ras and Ras-related GTPases specifically Cdc42 activated mutant, round 2Confirmatory same project related to Summary assay
588626SAR compounds tested by Multiplex dose response to identify specific small molecule inhibitors of Ras and Ras-related GTPases specifically Cdc42 wildtype, round 2Confirmatory same project related to Summary assay
588628SAR compounds tested by Multiplex dose response to identify specific small molecule inhibitors of Ras and Ras-related GTPases specifically H-Ras activated mutant, round 2Confirmatory same project related to Summary assay
588630SAR compounds tested by Multiplex dose response to identify specific small molecule inhibitors of Ras and Ras-related GTPases specifically H-Ras wildtype, round 2Confirmatory same project related to Summary assay
588631SAR compounds tested by Multiplex dose response to identify specific small molecule inhibitors of Ras and Ras-related GTPases specifically Rab7 wildtype, round 2Confirmatory same project related to Summary assay
602137Counter screen to determine effect of SAR compounds for Rab7 project on GST-GSH binding, round 1Other same project related to Summary assay
602145Counter screen to determine effect of SAR compounds for Rab7 project on GST-GSH binding, round 2Other same project related to Summary assay
602146EGFR degradation assay in SCC-12F cellsOther same project related to Summary assay
651732Cellular toxicity assessed by Trypan Blue for compounds in active Cdc42 Probe Extension ProjectOther same project related to Summary assay
652107Actin Polymerization inhibition by compounds from Cdc42 Probe Extension ProjectOther same project related to Summary assay
720688Assessing protein source dependence on dose response curves of small molecule inhibitors of Ras and Ras-related GTPases specifically Cdc42 wildtypeOther same project related to Summary assay
720689Assessing protein source dependence on dose response curves of small molecule inhibitors of Ras and Ras-related GTPases specifically Rab7 wildtypeOther same project related to Summary assay
720699Comparison of test compound incubation dependence on dose response curves of small molecule inhibitors of Ras and Ras-related GTPases specifically Rab7 wildtypeOther same project related to Summary assay
720712Comparison of test compound incubation dependence on dose response curves of small molecule inhibitors of Ras and Ras-related GTPases specifically Cdc42 wildtypeOther same project related to Summary assay
743330 On Hold
Description:
University of New Mexico Assay Overview:
Assay Support: NIH I RO3 MH081231-01
HTS to identify specific small molecule inhibitors of Ras and Ras-related GTPases
PI: Angela Wandinger-Ness, Ph.D.
Co-PI: Larry Sklar, Ph.D.
Assay Implementation: Peter Simons, Ph.D., Lin Hong, Ph.D.
Target Team Leader for the Center: Larry Sklar (lsklar@salud.unm.edu)
UNM Cheminformatics: Cristian Bologa, Ph.D., Oleg Ursu, Ph.D.

Chemistry: University of Kansas Specialized Chemistry Center
KU Specialized Chemistry Center PI: Jeff Aube, Ph.D.
KU SCC Project Manager: Jennifer E. Golden. Ph.D.
KU SCC Chemists on this project: Chad Schroeder, M.S., Denise Simpson, Ph.D., Julica Noeth, B.S.

Assay Background and Significance:
Activation of GTPases (in particular Rho family) are involved in the signal transduction cascades that commences with stimulation of formyl peptide receptors and resulting in the increase ligand-affinity of VLA-4 integrins. Assessment of this increased affinity can be made with , fluorescently labeled specific ligand for VLA-4, LDV-FITC. By utilizing U937 cells expressing non-desensetizing, deltaST, formyl peptide receptors, VLA-4 remains activate for a longer period of time post-stimulation. Hence a compound that interferes with the prolonged activation of GTPase would result in decreased ligand affinity.
Protocol
Cells (U937 cells expressing deltaST, non-desentizied formyl peptide receptors) are incubated at 37 degrees for 10-20 min. Flow cytometer data are acquired for 1024 s at 37 degrees. After establishing a baseline for unstained cells, LDV-FITC (VLA-4-specific ligand 4-((N'-2-methylphenyl)ureido)-phenylacetyl-l-leucyl-l-aspartyl-l-valyl-l-prolyl-l-alanyl-l-alanyl-l-lysine (LDV) FITC-conjugated) is added at sub-saturating concentration (~4 nanoM). Approximately 2 min later, fMLFF peptide (100 nanoM) is added to induce the cell stimulation. When the fluorescence increase stabilizes, different concentrations of compounds are added. The mean channel fluorescence is recorded. Nonspecific binding of LDV-FITC is determined by an excess of unlabeled LDV.
Response Assessment:
Compounds that induce a temporal decrease in LDV-FITC binding are considered to be active. Three distinct activity levels were assigned to each compounds; 0 no activity, 1 weak activity, and 2 strong activity.
PUBCHEM_ACTIVITY_SCORE is based on the different levels of activity:
Non-active compounds, SCORE = 0
Weak active compounds, SCORE = 33
Strong active compounds, SCORE = 66
Active compounds have SCORE > 0
Result Definitions
TIDNameDescriptionHistogramTypeUnit
OutcomeThe BioAssay activity outcomeOutcome
ScoreThe BioAssay activity ranking scoreInteger
1VLA4_Affinity_Level (50μM**)Level of change in VLA-4 affinity due to test compoundInteger

** Test Concentration.
Additional Information
Grant Number: NIH I RO3 MH081231-01

Data Table (Concise)
Data Table ( Complete ):     View Active Data    View All Data
Classification
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