Coupled Bioluminescent Assays: Methods, Evaluations, and Applications

315 Pages·2008·11.884 MB·English

by  Michael J. Corey(auth.)

COUPLED
BIOLUMINESCENT
ASSAYS
COUPLED
BIOLUMINESCENT
ASSAYS
Methods, Evaluations, and Applications
MICHAELJ.COREY
AJohnWiley&Sons,Inc.,Publication
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ISBN978-0-470-10883-3
PrintedintheUnitedStatesofAmerica
10987654321
“Therearetwowaysofspreadinglight:
tobethecandleorthemirrorthatreflectsit.”
–EdithWharton
CONTENTS
PREFACE xiii
PARTI BACKGROUNDTOCOUPLED
BIOLUMINESCENTASSAYS 1
1 Introduction 3
1.1 IntroductiontoCoupledBioluminescentAssays, 3
1.2 LuminescentTechnologiesoftheLifeSciences, 4
1.3 VarietiesofFluorometricAssays, 7
1.3.1 Time-ResolvedFluorescence, 8
1.3.2 FluorescenceResonanceEnergyTransfer, 8
1.3.3 FluorescencePolarization, 9
1.4 ChemiluminescenceandBioluminescence, 10
1.4.1 Chemiluminescence, 11
1.4.2 Bioluminescence, 13
1.5 CommonBioluminescenceSystems, 14
1.5.1 FireflyLuciferase, 14
1.5.2 RenillaLuciferase, 16
1.5.3 BacterialLuciferases, 18
1.5.4 Aequorin, 19
1.5.5 OtherLuciferasesandPhotoproteins, 20
1.6 ACoupledBioluminescentReaction, 22
1.7 Summary, 23
vii
viii CONTENTS
2 CoupledBioluminescentReactionsinPractice 24
2.1 PrinciplesofCoupledBioluminescentReactions, 24
2.1.1 RequirementsforSuccessfulCoupledBioluminescent
Assays, 24
2.2 InstrumentationandEquipmentforCoupledBioluminescent
Assays, 25
2.2.1 InstrumentTesting, 27
2.2.2 InstrumentFeatures, 28
2.2.3 LuminescentMicroplates, 31
2.3 CoupledBioluminescentAssayProceduresandPrecautions, 32
2.3.1 CoupledEnzymeAssays, 32
2.3.2 Steady-StateKinetics, 36
2.3.3 CoupledBioluminescentReactions, 38
2.3.4 RecommendedPrecautionsforCoupled
BioluminescentReactions, 44
2.4 DataHandlingforCoupledBioluminescentAssays, 45
2.4.1 AnalysisofCoupledBioluminescentAssayResults
byTime-DependentLinearRegression, 46
2.4.2 PerformingtheLinearRegression, 49
2.4.3 Outliers, 50
2.4.4 SpecialConsiderationsRelatingtoHigh-Throughput
Screening, 52
2.5 ComparisonofCoupledBioluminescentAssayswith
OtherMethods, 53
2.5.1 CoupledBioluminescentAssaysVersus
SpectrophotometricAssays, 53
2.5.2 CoupledBioluminescentAssaysVersus
FluorescentAssays, 54
2.5.3 CoupledBioluminescentAssays
VersusChemiluminescentAssays, 55
PARTII BIOMEDICALAPPLICATIONSOFCOUPLED
BIOLUMINESCENCE 57
3 CoupledBioluminescentCytotoxicityAssays 59
3.1 Introduction, 59
3.2 MembraneIntegrityAssays, 60
3.2.1 RadioactiveIsotopeMethods, 60
3.2.2 OtherLabelingMethods, 64
3.2.3 GeneralEnzyme-ReleaseMethods, 65
3.3 ACoupledBioluminescentAssayforEnzymeRelease:
TheG3PDH-ReleaseAssay, 70
3.3.1 DevelopmentandFeatures, 70
3.3.2 PerformanceCharacteristics, 75
CONTENTS ix
3.4 ViabilityAssays, 82
3.4.1 ATP-ReleaseAssay, 82
3.4.2 AssaysofVitalCellularFunctions, 82
3.4.3 SimpleCell-GrowthAssays, 83
3.4.4 TotalEnzyme-ReleaseMethods, 84
4 TheRoleofCoupledBioluminescentAssaysinKinaseScreening
andStudy 85
4.1 TheManyRolesofKinasesinBiology, 85
4.2 CurrentStandardKinaseAssays, 86
4.2.1 FluorometricKinaseAssays, 87
4.2.2 RadiolabelingKinaseAssays, 88
4.2.3 FluorometricAssaysInvolvingDetectionofADP, 89
4.3 CoupledBioluminescentKinaseAssays, 90
4.3.1 AHistoricalNote:TheCoupledBioluminescentCreatine
KinaseAssay, 90
4.3.2 ATPDepletionKinaseAssays, 90
4.3.3 ReverseCBKinaseAssays, 98
4.3.4 KinasesasParticipantsinOtherCBAssays, 102
4.4 Conclusions, 102
5 CoupledBioluminescentPhosphataseAssays 103
5.1 Introduction, 103
5.2 Phosphatases, 103
5.2.1 CatalyticPropertiesofPhosphatases, 104
5.2.2 ClassificationofPhosphatases, 104
5.3 ContemporaryPhosphataseAssayTechnologies, 105
5.3.1 FluorometricPhosphataseAssays, 106
5.3.2 ColorimetricPhosphataseAssays, 117
5.3.3 ChemiluminescentPhosphataseAssays, 119
5.3.4 RadiolabelingPhosphataseAssays, 120
5.4 CBPhosphataseAssays, 122
5.4.1 AssaysUsingPhosphorylatedLuciferin, 122
5.4.2 AssaysBasedontheGPLReactionSeries, 122
5.4.3 ChallengesintheDevelopmentoftheGPL-Coupled
BioluminescentSystemforPhosphateDetection, 127
5.4.4 PotentialDrawbacksofGPLPhosphataseAssays, 129
5.5 Conclusions, 130
6 Acetylcholinesterase 131
6.1 Introduction, 131
6.2 EstablishedAChEAssayMethods, 133
6.3 RecentDevelopmentsinAChEAssayMethods, 134
x CONTENTS
6.4 EvaluationofCurrentAChEHigh-ThroughputScreening
Methods, 135
6.5 CoupledBioluminescentAssaysofAChEActivity, 136
6.6 ComparisonofCoupledBioluminescentandOtherMethodsof
MeasuringAChEActivity, 138
7 MeasurementofNitricOxideSynthaseActivityby
CoupledBioluminescence 139
7.1 Introduction, 139
7.1.1 NitricOxideSynthases, 139
7.1.2 TheNitrateIoninMedicineandNOBiology, 141
7.1.3 NOBiology, 141
7.1.4 InterrelationshipAmongNitrate,Nitrite,andNO, 142
7.1.5 MedicalAspectsofNitrateBiology, 143
7.2 CurrentNOSAssays, 144
7.2.1 MeasuringNOSActivitybyAssaysforNitriteandNitrate, 144
7.2.2 “Direct”AssaysofNOSActivity, 148
7.2.3 CoupledEnzymaticAssaysofNOSActivity, 148
7.3 Conclusions, 149
8 TheCoupledBioluminescentPyrophosphorolysisAssay 150
8.1 Introduction, 150
8.2 GeneticVariationinModernMedicine, 150
8.3 DNAComplementarity, 152
8.3.1 DNAHybridization, 152
8.3.2 PrimerExtensioninSNPAnalysis, 153
8.3.3 PrimerDegradationinSNPAnalysis, 154
8.4 TheREADIT®PyrophosphorolysisAssay, 156
8.4.1 READIT®:DescriptionandTechnicalAspects, 156
8.4.2 PerformingtheREADIT®Assay, 157
8.4.3 CompetitivePositionofREADIT®, 158
9 CoupledLuminescentAssaysofG-Protein-CoupledReceptors 160
9.1 IntroductiontoG-Protein-CoupledReceptors, 160
9.1.1 StructureandFunctionofGPCRs, 161
9.1.2 GPCRsinMedicine, 165
9.2 GPCRAssayMethods, 166
9.2.1 ArrestinsasMarkersofGPCRActivity, 166
9.2.2 CalciumQuantificationinGPCRAssays, 169
9.2.3 QuantitativecAMPAssaysofGPCRActivity, 175
9.2.4 GPCRAssaysInvolvingInositolTriphosphateDetection, 182
9.2.5 GPCRAssaysInvolvingGTP, 183
9.2.6 GPCRAssaysInvolvingReporterGenes, 186
9.2.7 NovelGPCRAssaysandOtherStrategies, 189
9.3 Summary, 190
CONTENTS xi
10 CoupledBioluminescentProteaseAssays 191
10.1 Introduction, 191
10.1.1 Proteases, 191
10.2 ProteaseAssays, 193
10.2.1 ChromogenicProteaseAssays, 194
10.2.2 FluorometricProteaseAssays, 194
10.2.3 ExoticFluorometricProteaseAssays, 197
10.3 CoupledBioluminescentProteaseAssays, 198
10.3.1 ProteaseAssaysUsingLuciferinDerivatives, 198
10.3.2 CoupledBioluminescentProteaseAssaysEmploying
RecombinantLuminescentProteins, 198
10.4 Summary, 199
11 CoupledLuminescentAssaysInvolvingAequorin 201
11.1 IntroductiontoAequorin, 201
11.1.1 AequorinastheFirstPhotoprotein, 201
11.1.2 TheAequorinReaction, 202
11.2 DetectionofCalciuminPractice, 203
11.2.1 FluorometricQuantificationofCalcium, 203
11.2.2 CoupledBioluminescentQuantificationofCalcium
UsingAequorin, 207
11.2.3 Real-TimeImagingofCalciumFlux
withAequorin, 208
11.2.4 OtherUsesofAequorininCoupled
BioluminescentAssays, 208
12 CoupledBioluminescentReporterAssays 209
12.1 IntroductiontoReporterAssays, 209
12.1.1 BriefHistoryandDevelopment
ofReporterAssays, 210
12.1.2 OtherNon-LuminescentReporterProteins, 211
12.2 LuciferasesasReportersofPromoterActivities, 212
12.2.1 TheModernLuminescentReporterAssay, 213
12.2.2 MulticolorLightEmissionbyLuciferases, 214
12.2.3 Dual-ModeReporterAssaysinPractice, 214
12.3 AequorinasaReporterEnzyme, 215
12.4 VectorsforUseinReporterAssays, 216
12.4.1 SystemsforStudyofSpecificTranscriptional
Modulators, 216
12.4.2 BacterialReporterSystems, 216
12.4.3 ViralVectors, 217
12.5 Summary, 217
xii CONTENTS
13 CoupledBioluminescentAssays:RegulatoryConcerns 218
13.1 Introduction, 218
13.2 RegulatoryAspectsofAssayDevelopment, 220
13.2.1 StandardOperatingProcedures, 220
13.2.2 PhilosophyofAssayValidation, 221
13.2.3 ParametersforAssayValidation, 223
13.2.4 GainingRegulatoryAcceptanceofCBMethods, 240
13.3 Summary, 240
PARTIII OTHERAPPLICATIONS
OFCOUPLEDBIOLUMINESCENCE 241
14 CoupledBioluminescentDeterminationofBioburdenandSterility 243
14.1 Introduction, 243
14.2 RapidMethodsofBioburdenandSterilityAssessment, 244
14.2.1 BioburdenMeasurementbyATP-ReleaseAssay, 244
14.2.2 BioburdenMeasurementbyProteinAssay, 245
14.2.3 CoupledBioluminescentMethodsofMeasuring
Bioburden, 245
15 EnvironmentalApplicationsofCoupledBioluminescentAssays 247
15.1 Introduction, 247
15.2 CurrentMethodsforEnvironmentalMonitoring
ofWaterQuality, 248
15.3 MethodsofMonitoringStreamWaterandLakeWater, 249
15.3.1 MonitoringofPhosphateinFreshwater, 249
15.3.2 MonitoringofNitrateinGroundwater, 250
15.3.3 MonitoringofPesticidesinFreshwater, 251
15.3.4 MonitoringofBacterialContentofFreshwater, 252
15.4 MethodsofMonitoringDrinkingWater, 254
15.4.1 CurrentDrinkingWaterReports, 254
15.4.2 BiohazardMonitoringinDrinkingWater, 255
15.4.3 MonitoringofChemicalHazardsinDrinkingWater, 257
AppendixA:One-LetterAminoAcidAbbreviations 259
Glossary 260
Bibliography 265
Index 291