Details of experiments between P61981 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: 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: 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): 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: 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)

Measured dissociation constant (10-6M): <1
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: 6


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: 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: 0.6
Holdup BI standard deviation: 0.01
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): 9.49
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: 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: 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): 129
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 7.9
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: 6.14


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: 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: 0.83
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): 7.74
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: 4.59


Experiment series 6

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: 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: 0.49
Holdup BI standard deviation: 0.02
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): 7.51
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: 3.94


Experiment series 7

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: 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): 115
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: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 4
Measured pKd: 6.2


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: 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: 0.86
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): 5.13
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
Measured pKd: 4.76


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: 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: 0.58
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): 4.81
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: 4.02


Experiment series 10

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: 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: 0.84
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): 4.46
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: 4.6


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: 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: 0.57
Holdup BI standard deviation: 0.28
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): 1.87
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: 3.99


Experiment series 12

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: 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: 0.22
Holdup BI standard deviation: 0.14
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): 1.68
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 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: 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: 0.16
Holdup BI standard deviation: 0.14
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): 1.11
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: 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: 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)

Average holdup BI: 0.35
Holdup BI standard deviation: 0.18
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): 0.9
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: 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: -0.08
Holdup BI standard deviation: 0.1
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): 0.88
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
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 construct: avi-his6-MBP-TEVsite-14-3-3gamma

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: 0.08
Holdup BI standard deviation: 0.1
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): 0.77
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: 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: 0.33
Holdup BI standard deviation: 0.41
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): 0.75
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: 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: 0.08
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): 0.65
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 19

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: 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: -0.03
Holdup BI standard deviation: 0.04
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): 0.58
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
The affinity was below the detection threshold of the assay.


Experiment series 20

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: 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: 0.17
Holdup BI standard deviation: 0.14
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): 0.57
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 21

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: 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: 0.01
Holdup BI standard deviation: 0.03
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): 0.31
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
The affinity was below the detection threshold of the assay.


Experiment series 22

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: 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: 0.15
Holdup BI standard deviation: 0.42
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): 0.3
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 23

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: 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: -0.03
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): 0.24
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 24

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: 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: 0.04
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): 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: 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: 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: -0.39
Holdup BI standard deviation: 1.61
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): 0.21
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 26

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: 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: -0.39
Holdup BI standard deviation: 1.54
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): 0.2
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 27

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: 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: -0.1
Holdup BI standard deviation: 0.6
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 28

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: 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: -0.08
Holdup BI standard deviation: 0.6
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): 0.1
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 29

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: 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: 0.79
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): 0.09
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 30

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: 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: -0.01
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): 0.07
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 31

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: 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: 1.44
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): 0.04
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: 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: 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: -0.01
Holdup BI standard deviation: 0.45
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): 0.02
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.


Experiment series 33

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: 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): 0.43
Standard deviation of dissociation constant (10-6M): 0.06
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: 6.37