5W4S

Solution structure of C2 domain from protein kinase C alpha in ternary complex with calcium and V5-pHM peptide


SOLUTION NMR
NMR Experiment
ExperimentTypeSample ContentsSolventIonic StrengthpHPressureTemperature (K)Spectrometer
12D 1H-15N HSQC2.225 mM CALCIUM ION, 0.89 mM [U-13C; U-15N] C2 domain of protein kinase C alpha, 6.7 mM MES, 67 mM potassium chloride, 0.2 mg/mL sodium azide, 2 mM V5-pHM peptide92% H2O/8% D2O0.076 M6.01 atm296.15Bruker AVANCE III 800
22D 1H-15N HSQC2.225 mM CALCIUM ION, 0.89 mM [U-13C; U-15N] C2 domain of protein kinase C alpha, 6.7 mM MES, 67 mM potassium chloride, 0.2 mg/mL sodium azide, 2 mM V5-pHM peptide92% H2O/8% D2O0.076 M6.01 atm296.15Bruker AVANCE III 500
32D 1H-15N HSQC3.75 mM CALCIUM ION, 1.47 mM [U-13C; U-15N] C2 domain of protein kinase C alpha, 6.7 mM MES, 67 mM potassium chloride, 0.2 mg/mL sodium azide, 0.6 mM V5-pHM peptide92% H2O/8% D2O0.077 M6.01 atm296.15Bruker AVANCE III 500
42D 1H-13C HSQC aliphatic2.225 mM CALCIUM ION, 0.89 mM [U-13C; U-15N] C2 domain of protein kinase C alpha, 6.7 mM MES, 67 mM potassium chloride, 0.2 mg/mL sodium azide, 2 mM V5-pHM peptide92% H2O/8% D2O0.076 M6.01 atm296.15Bruker AVANCE III 500
52D 1H-13C HSQC aliphatic2.225 mM CALCIUM ION, 0.89 mM [U-13C; U-15N] C2 domain of protein kinase C alpha, 6.7 mM MES, 67 mM potassium chloride, 0.2 mg/mL sodium azide, 2 mM V5-pHM peptide100% D2O0.076 M6.01 atm296.15Bruker AVANCE III 800
62D 1H-13C HSQC aromatic2.225 mM CALCIUM ION, 0.89 mM [U-13C; U-15N] C2 domain of protein kinase C alpha, 6.7 mM MES, 67 mM potassium chloride, 0.2 mg/mL sodium azide, 2 mM V5-pHM peptide100% D2O0.076 M6.01 atm296.15Bruker AVANCE III 800
103D C(CO)NH2.225 mM CALCIUM ION, 0.89 mM [U-13C; U-15N] C2 domain of protein kinase C alpha, 6.7 mM MES, 67 mM potassium chloride, 0.2 mg/mL sodium azide, 2 mM V5-pHM peptide92% H2O/8% D2O0.076 M6.01 atm296.15Bruker AVANCE III 500
93D H(CCO)NH2.225 mM CALCIUM ION, 0.89 mM [U-13C; U-15N] C2 domain of protein kinase C alpha, 6.7 mM MES, 67 mM potassium chloride, 0.2 mg/mL sodium azide, 2 mM V5-pHM peptide92% H2O/8% D2O0.076 M6.01 atm296.15Bruker AVANCE III 500
83D HCCH-TOCSY2.225 mM CALCIUM ION, 0.89 mM [U-13C; U-15N] C2 domain of protein kinase C alpha, 6.7 mM MES, 67 mM potassium chloride, 0.2 mg/mL sodium azide, 2 mM V5-pHM peptide92% H2O/8% D2O0.076 M6.01 atm296.15Bruker AVANCE III 500
73D HCCH-COSY2.225 mM CALCIUM ION, 0.89 mM [U-13C; U-15N] C2 domain of protein kinase C alpha, 6.7 mM MES, 67 mM potassium chloride, 0.2 mg/mL sodium azide, 2 mM V5-pHM peptide100% D2O0.076 M6.01 atm296.15Bruker AVANCE III 800
113D aromatic HCCH-TOCSY2.225 mM CALCIUM ION, 0.89 mM [U-13C; U-15N] C2 domain of protein kinase C alpha, 6.7 mM MES, 67 mM potassium chloride, 0.2 mg/mL sodium azide, 2 mM V5-pHM peptide100% D2O0.076 M6.01 atm296.15Bruker AVANCE III 800
123D 1H-13C NOESY aliphatic2.225 mM CALCIUM ION, 0.89 mM [U-13C; U-15N] C2 domain of protein kinase C alpha, 6.7 mM MES, 67 mM potassium chloride, 0.2 mg/mL sodium azide, 2 mM V5-pHM peptide100% D2O0.076 M6.01 atm296.15Bruker AVANCE III 800
133D 1H-15N NOESY2.225 mM CALCIUM ION, 0.89 mM [U-13C; U-15N] C2 domain of protein kinase C alpha, 6.7 mM MES, 67 mM potassium chloride, 0.2 mg/mL sodium azide, 2 mM V5-pHM peptide92% H2O/8% D2O0.076 M6.01 atm296.15Bruker AVANCE III 800
142D [F2-15N,13C filtered] NOESY2.225 mM CALCIUM ION, 0.89 mM [U-13C; U-15N] C2 domain of protein kinase C alpha, 6.7 mM MES, 67 mM potassium chloride, 0.2 mg/mL sodium azide, 2 mM V5-pHM peptide92% H2O/8% D2O0.076 M6.01 atm296.15Bruker AVANCE III 500
152D [F2-15N,13C filtered] NOESY2.225 mM CALCIUM ION, 0.89 mM [U-13C; U-15N] C2 domain of protein kinase C alpha, 6.7 mM MES, 67 mM potassium chloride, 0.2 mg/mL sodium azide, 2 mM V5-pHM peptide100% D2O0.076 M6.01 atm296.15Bruker AVANCE III 500
162D [F1,F2-15N,13C filtered] NOESY2.225 mM CALCIUM ION, 0.89 mM [U-13C; U-15N] C2 domain of protein kinase C alpha, 6.7 mM MES, 67 mM potassium chloride, 0.2 mg/mL sodium azide, 2 mM V5-pHM peptide92% H2O/8% D2O0.076 M6.01 atm296.15Bruker AVANCE III 500
172D [F1,F2-15N,13C filtered] NOESY3.75 mM CALCIUM ION, 1.47 mM [U-13C; U-15N] C2 domain of protein kinase C alpha, 6.7 mM MES, 67 mM potassium chloride, 0.2 mg/mL sodium azide, 0.6 mM V5-pHM peptide92% H2O/8% D2O0.077 M6.01 atm296.15Bruker AVANCE III 500
182D [F1,F2-15N,13C filtered] TOCSY2.225 mM CALCIUM ION, 0.89 mM [U-13C; U-15N] C2 domain of protein kinase C alpha, 6.7 mM MES, 67 mM potassium chloride, 0.2 mg/mL sodium azide, 2 mM V5-pHM peptide92% H2O/8% D2O0.076 M6.01 atm296.15Bruker AVANCE III 500
192D [F1,F2-15N,13C filtered] TOCSY3.75 mM CALCIUM ION, 1.47 mM [U-13C; U-15N] C2 domain of protein kinase C alpha, 6.7 mM MES, 67 mM potassium chloride, 0.2 mg/mL sodium azide, 0.6 mM V5-pHM peptide92% H2O/8% D2O0.077 M6.01 atm296.15Bruker AVANCE III 500
203D [F1-15N,13C filtered] 1H-15N NOESY2.225 mM CALCIUM ION, 0.89 mM [U-13C; U-15N] C2 domain of protein kinase C alpha, 6.7 mM MES, 67 mM potassium chloride, 0.2 mg/mL sodium azide, 2 mM V5-pHM peptide92% H2O/8% D2O0.076 M6.01 atm296.15Bruker AVANCE III 800
213D [F1-15N,13C filtered] 1H-13C NOESY2.225 mM CALCIUM ION, 0.89 mM [U-13C; U-15N] C2 domain of protein kinase C alpha, 6.7 mM MES, 67 mM potassium chloride, 0.2 mg/mL sodium azide, 2 mM V5-pHM peptide100% D2O0.076 M6.01 atm296.15Bruker AVANCE III 800
NMR Spectrometer Information
SpectrometerManufacturerModelField Strength
1BrukerAVANCE III800
2BrukerAVANCE III600
3BrukerAVANCE III500
NMR Refinement
MethodDetailsSoftware
DGSA-distance geometry simulated annealingCNS
NMR Ensemble Information
Conformer Selection Criteriastructures with the lowest energy
Conformers Calculated Total Number400
Conformers Submitted Total Number20
Representative Model1 (lowest energy)
Computation: NMR Software
#ClassificationVersionSoftware NameAuthor
1collectionTopSpinBruker Biospin
2processingNMRPipeDelaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax
3chemical shift assignmentSparkyGoddard
4structure calculationARIALinge, O'Donoghue and Nilges
5refinementCNSBrunger, Adams, Clore, Gros, Nilges and Read
6structure calculationHADDOCKBonvin