Details of experiments between P61981 and Q4KWH8
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: PLCH1
Protein B fragment: 1625-1634
Protein B site: 14-3-3_pSer1630
Protein B modification: (SEP)1630
Protein B sequence: AVNRHSTGSY
Protein B construct: AVNRH(pSer)TGSY (crude synthetic peptide)
Measured dissociation constant (10-6M): <100
Immobilized partner concentration in holdup experiment (10-6M): 134
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 1
Measured pKd: 4
Experiment series 2
Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: PLCH1
Protein B fragment: 1489-1498
Protein B site: 14-3-3_pSer1494
Protein B modification: (SEP)1494
Protein B sequence: PCKSKSLGDL
Protein B construct: PCKSK(pSer)LGDL (crude synthetic peptide)
Measured dissociation constant (10-6M): <10
Immobilized partner concentration in holdup experiment (10-6M): 129
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 1
Measured pKd: 5
Experiment series 3
Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: PLCH1
Protein B fragment: 965-974
Protein B site: 14-3-3_pSer970
Protein B modification: (SEP)970
Protein B sequence: QARPVSMPVD
Protein B construct: QARPV(pSer)MPVD (crude synthetic peptide)
Average holdup BI: 0.84
Holdup BI standard deviation: 0.01
Immobilized partner concentration in holdup experiment (10-6M): 129
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 8.31
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 5
Measured pKd: 4.61
Experiment series 4
Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: PLCH1
Protein B fragment: 1515-1524
Protein B site: 14-3-3_pThr1520
Protein B modification: (TPO)1520
Protein B sequence: SKSFVTTGIR
Protein B construct: SKSFV(pThr)TGIR (crude synthetic peptide)
Average holdup BI: 0.71
Holdup BI standard deviation: 0.01
Immobilized partner concentration in holdup experiment (10-6M): 115
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 7.79
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 4
Measured pKd: 4.34
Experiment series 5
Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: PLCH1
Protein B fragment: 965-974
Protein B site: 14-3-3_pSer970
Protein B modification: (SEP)970
Protein B sequence: QARPVSMPVD
Protein B construct: QARPV(pSer)MPVD (crude synthetic peptide)
Average holdup BI: 0.84
Holdup BI standard deviation: 0.04
Immobilized partner concentration in holdup experiment (10-6M): 134
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 6.15
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 4
Measured pKd: 4.58
Experiment series 6
Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: PLCH1
Protein B fragment: 1515-1524
Protein B site: 14-3-3_pThr1520
Protein B modification: (TPO)1520
Protein B sequence: SKSFVTTGIR
Protein B construct: SKSFV(pThr)TGIR (crude synthetic peptide)
Average holdup BI: 0.56
Holdup BI standard deviation: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 134
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 5.69
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 4
Measured pKd: 3.98
Experiment series 7
Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: PLCH1
Protein B fragment: 1515-1524
Protein B site: 14-3-3_pThr1520
Protein B modification: (TPO)1520
Protein B sequence: SKSFVTTGIR
Protein B construct: SKSFV(pThr)TGIR (crude synthetic peptide)
Average holdup BI: 0.59
Holdup BI standard deviation: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 129
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 5.58
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 5
Measured pKd: 4.06
Experiment series 8
Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: PLCH1
Protein B fragment: 965-974
Protein B site: 14-3-3_pSer970
Protein B modification: (SEP)970
Protein B sequence: QARPVSMPVD
Protein B construct: QARPV(pSer)MPVD (crude synthetic peptide)
Average holdup BI: 0.75
Holdup BI standard deviation: 0.07
Immobilized partner concentration in holdup experiment (10-6M): 115
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 5.05
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 4
Measured pKd: 4.44
Experiment series 9
Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: PLCH1
Protein B fragment: 918-927
Protein B site: 14-3-3_pSer923
Protein B modification: (SEP)923
Protein B sequence: LRRTASAPAK
Protein B construct: LRRTA(pSer)APAK (crude synthetic peptide)
Average holdup BI: 0.15
Holdup BI standard deviation: 0.13
Immobilized partner concentration in holdup experiment (10-6M): 129
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 1.3
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 5
The affinity was below the detection threshold of the assay.
Experiment series 10
Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: PLCH1
Protein B fragment: 1315-1324
Protein B site: 14-3-3_pThr1320
Protein B modification: (TPO)1320
Protein B sequence: LPKSPTKGED
Protein B construct: LPKSP(pThr)KGED (crude synthetic peptide)
Average holdup BI: -0.05
Holdup BI standard deviation: 0.06
Immobilized partner concentration in holdup experiment (10-6M): 115
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.82
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 4
The affinity was below the detection threshold of the assay.
Experiment series 11
Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: PLCH1
Protein B fragment: 1315-1324
Protein B site: 14-3-3_pThr1320
Protein B modification: (TPO)1320
Protein B sequence: LPKSPTKGED
Protein B construct: LPKSP(pThr)KGED (crude synthetic peptide)
Average holdup BI: -0.03
Holdup BI standard deviation: 0.08
Immobilized partner concentration in holdup experiment (10-6M): 129
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.41
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 5
The affinity was below the detection threshold of the assay.
Experiment series 12
Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: PLCH1
Protein B fragment: 1411-1420
Protein B site: 14-3-3_pSer1416
Protein B modification: (SEP)1416
Protein B sequence: ETLRPSVPEI
Protein B construct: ETLRP(pSer)VPEI (crude synthetic peptide)
Average holdup BI: -0.04
Holdup BI standard deviation: 0.12
Immobilized partner concentration in holdup experiment (10-6M): 115
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.28
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 4
The affinity was below the detection threshold of the assay.
Experiment series 13
Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: PLCH1
Protein B fragment: 1315-1324
Protein B site: 14-3-3_pThr1320
Protein B modification: (TPO)1320
Protein B sequence: LPKSPTKGED
Protein B construct: LPKSP(pThr)KGED (crude synthetic peptide)
Average holdup BI: 0.04
Holdup BI standard deviation: 0.12
Immobilized partner concentration in holdup experiment (10-6M): 134
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.25
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 4
The affinity was below the detection threshold of the assay.
Experiment series 14
Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: PLCH1
Protein B fragment: 1489-1498
Protein B site: 14-3-3_pSer1494
Protein B modification: (SEP)1494
Protein B sequence: PCKSKSLGDL
Protein B construct: PCKSK(pSer)LGDL (crude synthetic peptide)
Average holdup BI: -0.49
Holdup BI standard deviation: 2.83
Immobilized partner concentration in holdup experiment (10-6M): 134
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.14
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 4
The affinity was below the detection threshold of the assay.
Experiment series 15
Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: PLCH1
Protein B fragment: 1411-1420
Protein B site: 14-3-3_pSer1416
Protein B modification: (SEP)1416
Protein B sequence: ETLRPSVPEI
Protein B construct: ETLRP(pSer)VPEI (crude synthetic peptide)
Average holdup BI: -0.03
Holdup BI standard deviation: 0.2
Immobilized partner concentration in holdup experiment (10-6M): 129
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.13
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 5
The affinity was below the detection threshold of the assay.
Experiment series 16
Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: PLCH1
Protein B fragment: 918-927
Protein B site: 14-3-3_pSer923
Protein B modification: (SEP)923
Protein B sequence: LRRTASAPAK
Protein B construct: LRRTA(pSer)APAK (crude synthetic peptide)
Average holdup BI: -0.02
Holdup BI standard deviation: 0.17
Immobilized partner concentration in holdup experiment (10-6M): 115
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.08
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 4
The affinity was below the detection threshold of the assay.
Experiment series 17
Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: PLCH1
Protein B fragment: 918-927
Protein B site: 14-3-3_pSer923
Protein B modification: (SEP)923
Protein B sequence: LRRTASAPAK
Protein B construct: LRRTA(pSer)APAK (crude synthetic peptide)
Average holdup BI: -0.01
Holdup BI standard deviation: 0.16
Immobilized partner concentration in holdup experiment (10-6M): 134
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.03
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 4
The affinity was below the detection threshold of the assay.
Experiment series 18
Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: PLCH1
Protein B fragment: 1515-1524
Protein B site: 14-3-3_pThr1520
Protein B modification: (TPO)1520
Protein B sequence: SKSFVTTGIR
Protein B construct: biotin-ttds-SKSFV(pThr)TGIR (HPLC-purified synthetic peptide)
Measured dissociation constant (10-6M): 159
Standard deviation of dissociation constant (10-6M): 21
Experiment method: competitive Fluorescence Polarization
Details of affinity fitting: Competitive binding equation in ProFit
Date of measurement: 2022.01.18
Number of measurements: 3
Measured pKd: 3.8
Experiment series 19
Protein A protein: YWHAG
Protein A fragment: 1-247
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: PLCH1
Protein B fragment: 1515-1524
Protein B site: 14-3-3_pThr1520
Protein B modification: (TPO)1520
Protein B sequence: SKSFVTTGIR
Protein B construct: biotin-ttds-SKSFV(pThr)TGIR (HPLC-purified synthetic peptide)
Measured dissociation constant (10-6M): 136
Standard deviation of dissociation constant (10-6M): 24
Experiment method: competitive Fluorescence Polarization
Details of affinity fitting: Competitive binding equation in ProFit
Date of measurement: 2022.01.10
Number of measurements: 3
Measured pKd: 3.87