Details of experiments between P31947 and O43379
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: WDR62
Protein B fragment: 1223-1232
Protein B site: 14-3-3_pSer1228
Protein B modification: (SEP)1228
Protein B sequence: ISRSISLGDS
Protein B construct: ISRSI(pSer)LGDS (crude synthetic peptide)
Average holdup BI: 0.54
Holdup BI standard deviation: 0.02
Immobilized partner concentration in holdup experiment (10-6M): 111
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 7.58
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: 4.02
Experiment series 2
Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: WDR62
Protein B fragment: 1223-1232
Protein B site: 14-3-3_pSer1228
Protein B modification: (SEP)1228
Protein B sequence: ISRSISLGDS
Protein B construct: ISRSI(pSer)LGDS (crude synthetic peptide)
Average holdup BI: 0.64
Holdup BI standard deviation: 0.04
Immobilized partner concentration in holdup experiment (10-6M): 142
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 7.55
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: 4.09
Experiment series 3
Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: WDR62
Protein B fragment: 1223-1232
Protein B site: 14-3-3_pSer1228
Protein B modification: (SEP)1228
Protein B sequence: ISRSISLGDS
Protein B construct: ISRSI(pSer)LGDS (crude synthetic peptide)
Average holdup BI: 0.57
Holdup BI standard deviation: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 113
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 6.58
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: 4.06
Experiment series 4
Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: WDR62
Protein B fragment: 1244-1253
Protein B site: 14-3-3_pSer1249
Protein B modification: (SEP)1249
Protein B sequence: LRRPSSVGEL
Protein B construct: LRRPS(pSer)VGEL (crude synthetic peptide)
Average holdup BI: 0.3
Holdup BI standard deviation: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 111
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 5.01
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: 3.58
Experiment series 5
Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: WDR62
Protein B fragment: 1244-1253
Protein B site: 14-3-3_pSer1249
Protein B modification: (SEP)1249
Protein B sequence: LRRPSSVGEL
Protein B construct: LRRPS(pSer)VGEL (crude synthetic peptide)
Average holdup BI: 0.29
Holdup BI standard deviation: 0.07
Immobilized partner concentration in holdup experiment (10-6M): 142
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 3.74
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: 3.46
Experiment series 6
Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: WDR62
Protein B fragment: 1244-1253
Protein B site: 14-3-3_pSer1249
Protein B modification: (SEP)1249
Protein B sequence: LRRPSSVGEL
Protein B construct: LRRPS(pSer)VGEL (crude synthetic peptide)
Average holdup BI: 0.13
Holdup BI standard deviation: 0.06
Immobilized partner concentration in holdup experiment (10-6M): 113
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 2.05
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
The affinity was below the detection threshold of the assay.
Experiment series 7
Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: WDR62
Protein B fragment: 1337-1346
Protein B site: 14-3-3_pSer1342
Protein B modification: (SEP)1342
Protein B sequence: SAFRPSLPAP
Protein B construct: SAFRP(pSer)LPAP (crude synthetic peptide)
Average holdup BI: 0.1
Holdup BI standard deviation: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 111
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 1.83
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 8
Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: WDR62
Protein B fragment: 44-53
Protein B site: 14-3-3_pSer49
Protein B modification: (SEP)49
Protein B sequence: RRTRLSTASE
Protein B construct: RRTRL(pSer)TASE (crude synthetic peptide)
Average holdup BI: 0.18
Holdup BI standard deviation: 0.15
Immobilized partner concentration in holdup experiment (10-6M): 142
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 1.23
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 9
Protein A protein: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: WDR62
Protein B fragment: 892-901
Protein B site: 14-3-3_pSer897
Protein B modification: (SEP)897
Protein B sequence: ESLENSILDS
Protein B construct: ESLEN(pSer)ILDS (crude synthetic peptide)
Average holdup BI: -0.21
Holdup BI standard deviation: 0.27
Immobilized partner concentration in holdup experiment (10-6M): 142
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 1.09
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: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: WDR62
Protein B fragment: 892-901
Protein B site: 14-3-3_pSer897
Protein B modification: (SEP)897
Protein B sequence: ESLENSILDS
Protein B construct: ESLEN(pSer)ILDS (crude synthetic peptide)
Average holdup BI: 0.13
Holdup BI standard deviation: 0.16
Immobilized partner concentration in holdup experiment (10-6M): 113
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.7
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: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: WDR62
Protein B fragment: 1129-1138
Protein B site: 14-3-3_pSer1134
Protein B modification: (SEP)1134
Protein B sequence: MDRGGSQPRA
Protein B construct: MDRGG(pSer)QPRA (crude synthetic peptide)
Average holdup BI: 0.18
Holdup BI standard deviation: 0.35
Immobilized partner concentration in holdup experiment (10-6M): 142
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.45
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: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: WDR62
Protein B fragment: 1337-1346
Protein B site: 14-3-3_pSer1342
Protein B modification: (SEP)1342
Protein B sequence: SAFRPSLPAP
Protein B construct: SAFRP(pSer)LPAP (crude synthetic peptide)
Average holdup BI: 0.02
Holdup BI standard deviation: 0.07
Immobilized partner concentration in holdup experiment (10-6M): 113
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.26
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: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: WDR62
Protein B fragment: 1129-1138
Protein B site: 14-3-3_pSer1134
Protein B modification: (SEP)1134
Protein B sequence: MDRGGSQPRA
Protein B construct: MDRGG(pSer)QPRA (crude synthetic peptide)
Average holdup BI: -0.17
Holdup BI standard deviation: 1.24
Immobilized partner concentration in holdup experiment (10-6M): 111
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.11
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: SFN
Protein A fragment: 1-248
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: WDR62
Protein B fragment: 1337-1346
Protein B site: 14-3-3_pSer1342
Protein B modification: (SEP)1342
Protein B sequence: SAFRPSLPAP
Protein B construct: SAFRP(pSer)LPAP (crude synthetic peptide)
Average holdup BI: -0.01
Holdup BI standard deviation: 0.09
Immobilized partner concentration in holdup experiment (10-6M): 142
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.11
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.