Details of experiments between P61981 and P40818
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 modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3gamma
Protein B protein: USP8
Protein B fragment: 712-724
Protein B site: 14-3-3_pSer718
Protein B modification: (SEP)718
Protein B sequence: LKRSYSSPDI
Protein B construct: KLKRSY(pSer)SPDITQ (crude synthetic peptide)
Measured dissociation constant (10-6M): <10
Immobilized partner concentration in holdup experiment (10-6M): 115
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: 5
Experiment series 2
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: USP8
Protein B fragment: 713-722
Protein B site: 14-3-3_pSer718
Protein B modification: (SEP)718
Protein B sequence: LKRSYSSPDI
Protein B construct: LKRSY(pSer)SPDI (crude synthetic peptide)
Measured dissociation constant (10-6M): <10
Immobilized partner concentration in holdup experiment (10-6M): 115
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: 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: USP8
Protein B fragment: 712-724
Protein B site: 14-3-3_pSer718
Protein B modification: (SEP)718
Protein B sequence: LKRSYSSPDI
Protein B construct: KLKRSY(pSer)SPDITQ (crude synthetic peptide)
Measured dissociation constant (10-6M): <10
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: 5
Experiment series 4
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: USP8
Protein B fragment: 713-722
Protein B site: 14-3-3_pSer718
Protein B modification: (SEP)718
Protein B sequence: LKRSYSSPDI
Protein B construct: LKRSY(pSer)SPDI (crude synthetic peptide)
Average holdup BI: 0.79
Holdup BI standard deviation: 0.02
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): 9.5
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: 4.46
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: USP8
Protein B fragment: 712-724
Protein B site: 14-3-3_pSer718
Protein B modification: (SEP)718
Protein B sequence: LKRSYSSPDI
Protein B construct: KLKRSY(pSer)SPDITQ (crude synthetic peptide)
Average holdup BI: 0.99
Holdup BI standard deviation: 0
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): 8.65
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: 5.81
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: USP8
Protein B fragment: 713-722
Protein B site: 14-3-3_pSer718
Protein B modification: (SEP)718
Protein B sequence: LKRSYSSPDI
Protein B construct: LKRSY(pSer)SPDI (crude synthetic peptide)
Average holdup BI: 0.82
Holdup BI standard deviation: 0.02
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): 7.98
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: 4.55
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: USP8
Protein B fragment: 636-645
Protein B site: 14-3-3_pSer641
Protein B modification: (SEP)641
Protein B sequence: LTRARSEEMG
Protein B construct: LTRAR(pSer)EEMG (crude synthetic peptide)
Average holdup BI: 0.62
Holdup BI standard deviation: 0.09
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): 5.35
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: 4.1
Experiment series 8
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: USP8
Protein B fragment: 686-695
Protein B site: 14-3-3_pSer691
Protein B modification: (SEP)691
Protein B sequence: SSAPPSTPPT
Protein B construct: SSAPP(pSer)TPPT (crude synthetic peptide)
Average holdup BI: -0.1
Holdup BI standard deviation: 0.11
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): 1
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 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: USP8
Protein B fragment: 686-695
Protein B site: 14-3-3_pSer691
Protein B modification: (SEP)691
Protein B sequence: SSAPPSTPPT
Protein B construct: SSAPP(pSer)TPPT (crude synthetic peptide)
Average holdup BI: -0.08
Holdup BI standard deviation: 0.12
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): 0.86
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: USP8
Protein B fragment: 686-695
Protein B site: 14-3-3_pSer691
Protein B modification: (SEP)691
Protein B sequence: SSAPPSTPPT
Protein B construct: SSAPP(pSer)TPPT (crude synthetic peptide)
Average holdup BI: -0.03
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): 0.3
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: USP8
Protein B fragment: 636-645
Protein B site: 14-3-3_pSer641
Protein B modification: (SEP)641
Protein B sequence: LTRARSEEMG
Protein B construct: LTRAR(pSer)EEMG (crude synthetic peptide)
Average holdup BI: -0.03
Holdup BI standard deviation: 0.82
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): 0.02
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.