Interaction Details


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
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: YAP1
Fragment: 100-109
Site: 14-3-3_pSer105
Modification: (SEP)105
Sequence: EPKSHSRQAS
Construct: EPKSH(pSer)RQAS (crude synthetic peptide)
Measured dissociation constant (10-6M): <100
Immobilized partner concentration (10-6M): 142
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 1
Measured pKd: 4
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 2

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: YAP1
Fragment: 144-153
Site: 14-3-3_pSer149
Modification: (SEP)149
Sequence: PTGVVSGPAA
Construct: PTGVV(pSer)GPAA (crude synthetic peptide)
Average holdup BI: 0.25
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 113
P value: 6.52
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.47
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))

Experiment series 3

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: YAP1
Fragment: 144-153
Site: 14-3-3_pSer149
Modification: (SEP)149
Sequence: PTGVVSGPAA
Construct: PTGVV(pSer)GPAA (crude synthetic peptide)
Average holdup BI: 0.24
Holdup BI standard deviation: 0.08
Immobilized partner concentration (10-6M): 142
P value: 3.16
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.35
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 4

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: YAP1
Fragment: 144-153
Site: 14-3-3_pSer149
Modification: (SEP)149
Sequence: PTGVVSGPAA
Construct: PTGVV(pSer)GPAA (crude synthetic peptide)
Average holdup BI: 0.2
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 111
P value: 4.79
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))

Experiment series 5

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: YAP1
Fragment: 271-280
Site: 14-3-3_pSer276
Modification: (SEP)276
Sequence: QRISQSAPVK
Construct: QRISQ(pSer)APVK (crude synthetic peptide)
Average holdup BI: 0.99
Holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 113
P value: 6.19
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 5.76
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))

Experiment series 6

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: YAP1
Fragment: 271-280
Site: 14-3-3_pSer276
Modification: (SEP)276
Sequence: QRISQSAPVK
Construct: QRISQ(pSer)APVK (crude synthetic peptide)
Average holdup BI: 0.99
Holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 142
P value: 10.88
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 5.69
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 7

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: YAP1
Fragment: 271-280
Site: 14-3-3_pSer276
Modification: (SEP)276
Sequence: QRISQSAPVK
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
DOI: 10.1101/2022.12.08.519103

Experiment series 8

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: YAP1
Fragment: 271-280
Site: 14-3-3_pSer276
Modification: (SEP)276
Sequence: QRISQSAPVK
Construct: QRISQ(pSer)APVK (crude synthetic peptide)
Average holdup BI: 0.99
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 111
P value: 5.31
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 5.79
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))

Experiment series 9

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: YAP1
Fragment: 366-375
Site: 14-3-3_pSer371
Modification: (SEP)371
Sequence: SSPGMSQELR
Construct: SSPGM(pSer)QELR (crude synthetic peptide)
Average holdup BI: 0.15
Holdup BI standard deviation: 0.13
Immobilized partner concentration (10-6M): 113
P value: 1.16
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))

Experiment series 10

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: YAP1
Fragment: 366-375
Site: 14-3-3_pSer371
Modification: (SEP)371
Sequence: SSPGMSQELR
Construct: SSPGM(pSer)QELR (crude synthetic peptide)
Average holdup BI: 0.1
Holdup BI standard deviation: 0.14
Immobilized partner concentration (10-6M): 142
P value: 0.73
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 11

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: YAP1
Fragment: 366-375
Site: 14-3-3_pSer371
Modification: (SEP)371
Sequence: SSPGMSQELR
Construct: SSPGM(pSer)QELR (crude synthetic peptide)
Average holdup BI: -0.08
Holdup BI standard deviation: 0.28
Immobilized partner concentration (10-6M): 111
P value: 0.22
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))

Experiment series 12

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: YAP1
Fragment: 376-385
Site: 14-3-3_pSer381
Modification: (SEP)381
Sequence: TMTTNSSDPF
Construct: TMTTN(pSer)SDPF (crude synthetic peptide)
Average holdup BI: 0.09
Holdup BI standard deviation: 0.14
Immobilized partner concentration (10-6M): 142
P value: 0.58
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 13

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: YAP1
Fragment: 379-388
Site: 14-3-3_pSer384
Modification: (SEP)384
Sequence: DESTDSGLSM
Construct: DESTD(pSer)GLSM (crude synthetic peptide)
Average holdup BI: 0.09
Holdup BI standard deviation: 0.37
Immobilized partner concentration (10-6M): 113
P value: 0.13
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))

Experiment series 14

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: YAP1
Fragment: 382-391
Site: 14-3-3_pSer387
Modification: (SEP)387
Sequence: TDSGLSMSSY
Construct: TDSGL(pSer)MSSY (crude synthetic peptide)
Average holdup BI: -0.01
Holdup BI standard deviation: 0.11
Immobilized partner concentration (10-6M): 142
P value: 0.11
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 15

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: YAP1
Fragment: 383-392
Site: 14-3-3_pSer388
Modification: (SEP)388
Sequence: DPFLNSGTYH
Construct: DPFLN(pSer)GTYH (crude synthetic peptide)
Average holdup BI: -0.05
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 113
P value: 1.09
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))

Experiment series 16

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: YAP1
Fragment: 383-392
Site: 14-3-3_pSer388
Modification: (SEP)388
Sequence: DPFLNSGTYH
Construct: DPFLN(pSer)GTYH (crude synthetic peptide)
Average holdup BI: -0.05
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 142
P value: 0.8
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 17

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: YAP1
Fragment: 383-392
Site: 14-3-3_pSer388
Modification: (SEP)388
Sequence: DPFLNSGTYH
Construct: DPFLN(pSer)GTYH (crude synthetic peptide)
Average holdup BI: -0.06
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 111
P value: 0.82
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))

Experiment series 18

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: YAP1
Fragment: 56-65
Site: 14-3-3_pSer61
Modification: (SEP)61
Sequence: HVRGDSETDL
Construct: HVRGD(pSer)ETDL (crude synthetic peptide)
Average holdup BI: 0.35
Holdup BI standard deviation: 0.25
Immobilized partner concentration (10-6M): 113
P value: 1.38
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.68
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))

Experiment series 19

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: YAP1
Fragment: 56-65
Site: 14-3-3_pSer61
Modification: (SEP)61
Sequence: HVRGDSETDL
Construct: HVRGD(pSer)ETDL (crude synthetic peptide)
Average holdup BI: 0.14
Holdup BI standard deviation: 0.48
Immobilized partner concentration (10-6M): 142
P value: 0.23
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 20

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: YAP1
Fragment: 56-65
Site: 14-3-3_pSer61
Modification: (SEP)61
Sequence: HVRGDSETDL
Construct: HVRGD(pSer)ETDL (crude synthetic peptide)
Average holdup BI: 0.11
Holdup BI standard deviation: 0.21
Immobilized partner concentration (10-6M): 111
P value: 0.47
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))

Experiment series 21

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: YAP1
Fragment: 89-98
Site: 14-3-3_pSer94
Modification: (SEP)94
Sequence: RKLPDSFFKP
Construct: RKLPD(pSer)FFKP (crude synthetic peptide)
Average holdup BI: -0.09
Holdup BI standard deviation: 0.69
Immobilized partner concentration (10-6M): 142
P value: 0.08
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 22

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: YAP1
Fragment: 89-98
Site: 14-3-3_pSer94
Modification: (SEP)94
Sequence: RKLPDSFFKP
Construct: RKLPD(pSer)FFKP (crude synthetic peptide)
Average holdup BI: 0
Holdup BI standard deviation: 0.95
Immobilized partner concentration (10-6M): 111
P value: 0
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))

Experiment series 23

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: YAP1
Fragment: 105-114
Site: 14-3-3_pThr110
Modification: (TPO)110
Sequence: SRQASTDAGT
Construct: SRQAS(pThr)DAGT (crude synthetic peptide)
Measured dissociation constant (10-6M): <100
Immobilized partner concentration (10-6M): 113
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 1
Measured pKd: 4
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))

Experiment series 24

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: YAP1
Fragment: 109-118
Site: 14-3-3_pThr114
Modification: (TPO)114
Sequence: STDAGTAGAL
Construct: STDAG(pThr)AGAL (crude synthetic peptide)
Average holdup BI: -0.09
Holdup BI standard deviation: 0.35
Immobilized partner concentration (10-6M): 113
P value: 0.21
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))

Experiment series 25

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: YAP1
Fragment: 109-118
Site: 14-3-3_pThr114
Modification: (TPO)114
Sequence: STDAGTAGAL
Construct: STDAG(pThr)AGAL (crude synthetic peptide)
Average holdup BI: -0.14
Holdup BI standard deviation: 1.59
Immobilized partner concentration (10-6M): 111
P value: 0.07
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))

Experiment series 26

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: YAP1
Fragment: 136-145
Site: 14-3-3_pThr141
Modification: (TPO)141
Sequence: AVSPGTLTPT
Construct: AVSPG(pThr)LTPT (crude synthetic peptide)
Average holdup BI: -0.21
Holdup BI standard deviation: 0.57
Immobilized partner concentration (10-6M): 113
P value: 0.36
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))

Experiment series 27

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: YAP1
Fragment: 136-145
Site: 14-3-3_pThr141
Modification: (TPO)141
Sequence: AVSPGTLTPT
Construct: AVSPG(pThr)LTPT (crude synthetic peptide)
Average holdup BI: -0.11
Holdup BI standard deviation: 0.29
Immobilized partner concentration (10-6M): 111
P value: 0.33
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))

Experiment series 28

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: YAP1
Fragment: 140-149
Site: 14-3-3_pThr145
Modification: (TPO)145
Sequence: GTLTPTGVVS
Construct: GTLTP(pThr)GVVS (crude synthetic peptide)
Average holdup BI: 0.06
Holdup BI standard deviation: 0.1
Immobilized partner concentration (10-6M): 113
P value: 0.57
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))

Experiment series 29

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: YAP1
Fragment: 140-149
Site: 14-3-3_pThr145
Modification: (TPO)145
Sequence: GTLTPTGVVS
Construct: GTLTP(pThr)GVVS (crude synthetic peptide)
Average holdup BI: 0
Holdup BI standard deviation: 0.13
Immobilized partner concentration (10-6M): 142
P value: 0
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 30

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: YAP1
Fragment: 140-149
Site: 14-3-3_pThr145
Modification: (TPO)145
Sequence: GTLTPTGVVS
Construct: GTLTP(pThr)GVVS (crude synthetic peptide)
Average holdup BI: -0.06
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 111
P value: 1.19
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))

Experiment series 31

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: YAP1
Fragment: 72-81
Site: 14-3-3_pThr77
Modification: (TPO)77
Sequence: VMNPKTANVP
Construct: VMNPK(pThr)ANVP (crude synthetic peptide)
Average holdup BI: -0.06
Holdup BI standard deviation: 0.19
Immobilized partner concentration (10-6M): 142
P value: 0.23
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 32

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: YAP1
Fragment: 78-87
Site: 14-3-3_pThr83
Modification: (TPO)83
Sequence: ANVPQTVPMR
Construct: ANVPQ(pThr)VPMR (crude synthetic peptide)
Average holdup BI: 0.65
Holdup BI standard deviation: 0.1
Immobilized partner concentration (10-6M): 113
P value: 3.58
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.21
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))

Experiment series 33

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: YAP1
Fragment: 78-87
Site: 14-3-3_pThr83
Modification: (TPO)83
Sequence: ANVPQTVPMR
Construct: ANVPQ(pThr)VPMR (crude synthetic peptide)
Average holdup BI: 0.67
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 142
P value: 6.62
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.15
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 34

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: YAP1
Fragment: 78-87
Site: 14-3-3_pThr83
Modification: (TPO)83
Sequence: ANVPQTVPMR
Construct: ANVPQ(pThr)VPMR (crude synthetic peptide)
Average holdup BI: 0.64
Holdup BI standard deviation: 0.13
Immobilized partner concentration (10-6M): 111
P value: 3.73
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.2
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))