Details of experiments between P31947 and P46937
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 construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: YAP1
Protein B fragment: 105-114
Protein B site: 14-3-3_pThr110
Protein B modification: (TPO)110
Protein B sequence: SRQASTDAGT
Protein B construct: SRQAS(pThr)DAGT (crude synthetic peptide)
Measured dissociation constant (10-6M): <100
Immobilized partner concentration in holdup experiment (10-6M): 113
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: 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: YAP1
Protein B fragment: 100-109
Protein B site: 14-3-3_pSer105
Protein B modification: (SEP)105
Protein B sequence: EPKSHSRQAS
Protein B construct: EPKSH(pSer)RQAS (crude synthetic peptide)
Measured dissociation constant (10-6M): <100
Immobilized partner concentration in holdup experiment (10-6M): 142
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: 4
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: YAP1
Protein B fragment: 271-280
Protein B site: 14-3-3_pSer276
Protein B modification: (SEP)276
Protein B sequence: QRISQSAPVK
Protein B construct: QRISQ(pSer)APVK (crude synthetic peptide)
Average holdup BI: 0.99
Holdup BI standard deviation: 0
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): 10.88
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: 5.69
Experiment series 4
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: YAP1
Protein B fragment: 78-87
Protein B site: 14-3-3_pThr83
Protein B modification: (TPO)83
Protein B sequence: ANVPQTVPMR
Protein B construct: ANVPQ(pThr)VPMR (crude synthetic peptide)
Average holdup BI: 0.67
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): 6.62
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: 4.15
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: YAP1
Protein B fragment: 144-153
Protein B site: 14-3-3_pSer149
Protein B modification: (SEP)149
Protein B sequence: PTGVVSGPAA
Protein B construct: PTGVV(pSer)GPAA (crude synthetic peptide)
Average holdup BI: 0.25
Holdup BI standard deviation: 0.02
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): 6.52
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: 3.47
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: YAP1
Protein B fragment: 271-280
Protein B site: 14-3-3_pSer276
Protein B modification: (SEP)276
Protein B sequence: QRISQSAPVK
Protein B construct: QRISQ(pSer)APVK (crude synthetic peptide)
Average holdup BI: 0.99
Holdup BI standard deviation: 0
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): 6.19
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: 5.76
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: YAP1
Protein B fragment: 271-280
Protein B site: 14-3-3_pSer276
Protein B modification: (SEP)276
Protein B sequence: QRISQSAPVK
Protein B construct: QRISQ(pSer)APVK (crude synthetic peptide)
Average holdup BI: 0.99
Holdup BI standard deviation: 0.01
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): 5.31
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: 5.79
Experiment series 8
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: YAP1
Protein B fragment: 144-153
Protein B site: 14-3-3_pSer149
Protein B modification: (SEP)149
Protein B sequence: PTGVVSGPAA
Protein B construct: PTGVV(pSer)GPAA (crude synthetic peptide)
Average holdup BI: 0.2
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): 4.79
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
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 modification: 100_uM_FCA
Protein A construct: avi-his6-MBP-TEVsite-14-3-3sigma
Protein B protein: YAP1
Protein B fragment: 78-87
Protein B site: 14-3-3_pThr83
Protein B modification: (TPO)83
Protein B sequence: ANVPQTVPMR
Protein B construct: ANVPQ(pThr)VPMR (crude synthetic peptide)
Average holdup BI: 0.64
Holdup BI standard deviation: 0.13
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): 3.73
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: 4.2
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: YAP1
Protein B fragment: 78-87
Protein B site: 14-3-3_pThr83
Protein B modification: (TPO)83
Protein B sequence: ANVPQTVPMR
Protein B construct: ANVPQ(pThr)VPMR (crude synthetic peptide)
Average holdup BI: 0.65
Holdup BI standard deviation: 0.1
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): 3.58
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: 4.21
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: YAP1
Protein B fragment: 144-153
Protein B site: 14-3-3_pSer149
Protein B modification: (SEP)149
Protein B sequence: PTGVVSGPAA
Protein B construct: PTGVV(pSer)GPAA (crude synthetic peptide)
Average holdup BI: 0.24
Holdup BI standard deviation: 0.08
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): 3.16
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: 3.35
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: YAP1
Protein B fragment: 56-65
Protein B site: 14-3-3_pSer61
Protein B modification: (SEP)61
Protein B sequence: HVRGDSETDL
Protein B construct: HVRGD(pSer)ETDL (crude synthetic peptide)
Average holdup BI: 0.35
Holdup BI standard deviation: 0.25
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): 1.38
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: 3.68
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: YAP1
Protein B fragment: 140-149
Protein B site: 14-3-3_pThr145
Protein B modification: (TPO)145
Protein B sequence: GTLTPTGVVS
Protein B construct: GTLTP(pThr)GVVS (crude synthetic peptide)
Average holdup BI: -0.06
Holdup BI standard deviation: 0.06
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): 1.19
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: YAP1
Protein B fragment: 366-375
Protein B site: 14-3-3_pSer371
Protein B modification: (SEP)371
Protein B sequence: SSPGMSQELR
Protein B construct: SSPGM(pSer)QELR (crude synthetic peptide)
Average holdup BI: 0.15
Holdup BI standard deviation: 0.13
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): 1.16
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: 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: YAP1
Protein B fragment: 383-392
Protein B site: 14-3-3_pSer388
Protein B modification: (SEP)388
Protein B sequence: DPFLNSGTYH
Protein B construct: DPFLN(pSer)GTYH (crude synthetic peptide)
Average holdup BI: -0.05
Holdup BI standard deviation: 0.05
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): 1.09
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 16
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: YAP1
Protein B fragment: 383-392
Protein B site: 14-3-3_pSer388
Protein B modification: (SEP)388
Protein B sequence: DPFLNSGTYH
Protein B construct: DPFLN(pSer)GTYH (crude synthetic peptide)
Average holdup BI: -0.06
Holdup BI standard deviation: 0.07
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): 0.82
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 17
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: YAP1
Protein B fragment: 383-392
Protein B site: 14-3-3_pSer388
Protein B modification: (SEP)388
Protein B sequence: DPFLNSGTYH
Protein B construct: DPFLN(pSer)GTYH (crude synthetic peptide)
Average holdup BI: -0.05
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): 0.8
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 18
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: YAP1
Protein B fragment: 366-375
Protein B site: 14-3-3_pSer371
Protein B modification: (SEP)371
Protein B sequence: SSPGMSQELR
Protein B construct: SSPGM(pSer)QELR (crude synthetic peptide)
Average holdup BI: 0.1
Holdup BI standard deviation: 0.14
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): 0.73
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 19
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: YAP1
Protein B fragment: 376-385
Protein B site: 14-3-3_pSer381
Protein B modification: (SEP)381
Protein B sequence: TMTTNSSDPF
Protein B construct: TMTTN(pSer)SDPF (crude synthetic peptide)
Average holdup BI: 0.09
Holdup BI standard deviation: 0.14
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): 0.58
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
The affinity was below the detection threshold of the assay.
Experiment series 20
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: YAP1
Protein B fragment: 140-149
Protein B site: 14-3-3_pThr145
Protein B modification: (TPO)145
Protein B sequence: GTLTPTGVVS
Protein B construct: GTLTP(pThr)GVVS (crude synthetic peptide)
Average holdup BI: 0.06
Holdup BI standard deviation: 0.1
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): 0.57
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 21
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: YAP1
Protein B fragment: 56-65
Protein B site: 14-3-3_pSer61
Protein B modification: (SEP)61
Protein B sequence: HVRGDSETDL
Protein B construct: HVRGD(pSer)ETDL (crude synthetic peptide)
Average holdup BI: 0.11
Holdup BI standard deviation: 0.21
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): 0.47
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 22
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: YAP1
Protein B fragment: 136-145
Protein B site: 14-3-3_pThr141
Protein B modification: (TPO)141
Protein B sequence: AVSPGTLTPT
Protein B construct: AVSPG(pThr)LTPT (crude synthetic peptide)
Average holdup BI: -0.21
Holdup BI standard deviation: 0.57
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): 0.36
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 23
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: YAP1
Protein B fragment: 136-145
Protein B site: 14-3-3_pThr141
Protein B modification: (TPO)141
Protein B sequence: AVSPGTLTPT
Protein B construct: AVSPG(pThr)LTPT (crude synthetic peptide)
Average holdup BI: -0.11
Holdup BI standard deviation: 0.29
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): 0.33
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 24
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: YAP1
Protein B fragment: 72-81
Protein B site: 14-3-3_pThr77
Protein B modification: (TPO)77
Protein B sequence: VMNPKTANVP
Protein B construct: VMNPK(pThr)ANVP (crude synthetic peptide)
Average holdup BI: -0.06
Holdup BI standard deviation: 0.19
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): 0.23
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 25
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: YAP1
Protein B fragment: 56-65
Protein B site: 14-3-3_pSer61
Protein B modification: (SEP)61
Protein B sequence: HVRGDSETDL
Protein B construct: HVRGD(pSer)ETDL (crude synthetic peptide)
Average holdup BI: 0.14
Holdup BI standard deviation: 0.48
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): 0.23
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 26
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: YAP1
Protein B fragment: 366-375
Protein B site: 14-3-3_pSer371
Protein B modification: (SEP)371
Protein B sequence: SSPGMSQELR
Protein B construct: SSPGM(pSer)QELR (crude synthetic peptide)
Average holdup BI: -0.08
Holdup BI standard deviation: 0.28
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): 0.22
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 27
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: YAP1
Protein B fragment: 109-118
Protein B site: 14-3-3_pThr114
Protein B modification: (TPO)114
Protein B sequence: STDAGTAGAL
Protein B construct: STDAG(pThr)AGAL (crude synthetic peptide)
Average holdup BI: -0.09
Holdup BI standard deviation: 0.35
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): 0.21
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 28
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: YAP1
Protein B fragment: 379-388
Protein B site: 14-3-3_pSer384
Protein B modification: (SEP)384
Protein B sequence: DESTDSGLSM
Protein B construct: DESTD(pSer)GLSM (crude synthetic peptide)
Average holdup BI: 0.09
Holdup BI standard deviation: 0.37
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): 0.13
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 29
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: YAP1
Protein B fragment: 382-391
Protein B site: 14-3-3_pSer387
Protein B modification: (SEP)387
Protein B sequence: TDSGLSMSSY
Protein B construct: TDSGL(pSer)MSSY (crude synthetic peptide)
Average holdup BI: -0.01
Holdup BI standard deviation: 0.11
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.
Experiment series 30
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: YAP1
Protein B fragment: 89-98
Protein B site: 14-3-3_pSer94
Protein B modification: (SEP)94
Protein B sequence: RKLPDSFFKP
Protein B construct: RKLPD(pSer)FFKP (crude synthetic peptide)
Average holdup BI: -0.09
Holdup BI standard deviation: 0.69
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): 0.08
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 31
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: YAP1
Protein B fragment: 109-118
Protein B site: 14-3-3_pThr114
Protein B modification: (TPO)114
Protein B sequence: STDAGTAGAL
Protein B construct: STDAG(pThr)AGAL (crude synthetic peptide)
Average holdup BI: -0.14
Holdup BI standard deviation: 1.59
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): 0.07
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 32
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: YAP1
Protein B fragment: 89-98
Protein B site: 14-3-3_pSer94
Protein B modification: (SEP)94
Protein B sequence: RKLPDSFFKP
Protein B construct: RKLPD(pSer)FFKP (crude synthetic peptide)
Average holdup BI: 0
Holdup BI standard deviation: 0.95
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
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 33
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: YAP1
Protein B fragment: 140-149
Protein B site: 14-3-3_pThr145
Protein B modification: (TPO)145
Protein B sequence: GTLTPTGVVS
Protein B construct: GTLTP(pThr)GVVS (crude synthetic peptide)
Average holdup BI: 0
Holdup BI standard deviation: 0.13
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
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 34
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: YAP1
Protein B fragment: 271-280
Protein B site: 14-3-3_pSer276
Protein B modification: (SEP)276
Protein B sequence: QRISQSAPVK
Protein B construct: biotin-ttds-QRISQ(pSer)APVK (HPLC-purified synthetic peptide)
Measured dissociation constant (10-6M): 3.7
Standard deviation of dissociation constant (10-6M): 0.7
Experiment method: competitive Fluorescence Polarization
Details of affinity fitting: Competitive binding equation in ProFit
Date of measurement: 2022.02.09
Number of measurements: 3
Measured pKd: 5.43