Interaction Details


Details of experiments between P46937 and P27348
Note that the results of all experiments are listed, regardless of the modification states of the fragments.



Experiment series 1

Protein A

Protein: YAP1
Fragment: 144-153
Site: 14-3-3_pSer149
Modification: (SEP)149
Sequence: PTGVVSGPAA
Construct: PTGVV(pSer)GPAA (crude synthetic peptide)

Protein B

Protein: YWHAQ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3tau
Average holdup BI: 0.39
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 130
P value: 4.84
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.69
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: YAP1
Fragment: 271-280
Site: 14-3-3_pSer276
Modification: (SEP)276
Sequence: QRISQSAPVK
Construct: QRISQ(pSer)APVK (crude synthetic peptide)

Protein B

Protein: YWHAQ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3tau
Average holdup BI: 0.99
Holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 130
P value: 10.85
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 5.89
DOI: 10.1101/2022.12.08.519103
Experimental Details

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


Experiment series 3

Protein A

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)

Protein B

Protein: YWHAQ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3tau
Measured dissociation constant (10-6M): 1.8
Standard deviation of dissociation constant (10-6M): 0.17
Experiment method: competitive Fluorescence Polarization
Details of affinity fitting: Competitive binding equation in ProFit
Date of measurement: 2022-02-08
Number of measurements: 3
Measured pKd: 5.74
DOI: 10.1101/2022.12.08.519103

Experiment series 4

Protein A

Protein: YAP1
Fragment: 366-375
Site: 14-3-3_pSer371
Modification: (SEP)371
Sequence: SSPGMSQELR
Construct: SSPGM(pSer)QELR (crude synthetic peptide)

Protein B

Protein: YWHAQ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3tau
Average holdup BI: 0.05
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 130
P value: 2.18
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 5

Protein A

Protein: YAP1
Fragment: 376-385
Site: 14-3-3_pSer381
Modification: (SEP)381
Sequence: TMTTNSSDPF
Construct: TMTTN(pSer)SDPF (crude synthetic peptide)

Protein B

Protein: YWHAQ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3tau
Average holdup BI: 0.13
Holdup BI standard deviation: 0.14
Immobilized partner concentration (10-6M): 130
P value: 0.99
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 6

Protein A

Protein: YAP1
Fragment: 382-391
Site: 14-3-3_pSer387
Modification: (SEP)387
Sequence: TDSGLSMSSY
Construct: TDSGL(pSer)MSSY (crude synthetic peptide)

Protein B

Protein: YWHAQ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3tau
Average holdup BI: 0
Holdup BI standard deviation: 0.1
Immobilized partner concentration (10-6M): 130
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 7

Protein A

Protein: YAP1
Fragment: 383-392
Site: 14-3-3_pSer388
Modification: (SEP)388
Sequence: DPFLNSGTYH
Construct: DPFLN(pSer)GTYH (crude synthetic peptide)

Protein B

Protein: YWHAQ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3tau
Average holdup BI: -0.02
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 130
P value: 0.34
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 8

Protein A

Protein: YAP1
Fragment: 56-65
Site: 14-3-3_pSer61
Modification: (SEP)61
Sequence: HVRGDSETDL
Construct: HVRGD(pSer)ETDL (crude synthetic peptide)

Protein B

Protein: YWHAQ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3tau
Average holdup BI: 0.01
Holdup BI standard deviation: 0.48
Immobilized partner concentration (10-6M): 130
P value: 0.01
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 9

Protein A

Protein: YAP1
Fragment: 89-98
Site: 14-3-3_pSer94
Modification: (SEP)94
Sequence: RKLPDSFFKP
Construct: RKLPD(pSer)FFKP (crude synthetic peptide)

Protein B

Protein: YWHAQ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3tau
Average holdup BI: -0.13
Holdup BI standard deviation: 0.62
Immobilized partner concentration (10-6M): 130
P value: 0.18
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 10

Protein A

Protein: YAP1
Fragment: 109-118
Site: 14-3-3_pThr114
Modification: (TPO)114
Sequence: STDAGTAGAL
Construct: STDAG(pThr)AGAL (crude synthetic peptide)

Protein B

Protein: YWHAQ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3tau
Average holdup BI: 0.12
Holdup BI standard deviation: 1.02
Immobilized partner concentration (10-6M): 130
P value: 0.09
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: YAP1
Fragment: 140-149
Site: 14-3-3_pThr145
Modification: (TPO)145
Sequence: GTLTPTGVVS
Construct: GTLTP(pThr)GVVS (crude synthetic peptide)

Protein B

Protein: YWHAQ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3tau
Average holdup BI: 0.02
Holdup BI standard deviation: 0.24
Immobilized partner concentration (10-6M): 130
P value: 0.07
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 12

Protein A

Protein: YAP1
Fragment: 72-81
Site: 14-3-3_pThr77
Modification: (TPO)77
Sequence: VMNPKTANVP
Construct: VMNPK(pThr)ANVP (crude synthetic peptide)

Protein B

Protein: YWHAQ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3tau
Average holdup BI: -0.08
Holdup BI standard deviation: 0.38
Immobilized partner concentration (10-6M): 130
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 13

Protein A

Protein: YAP1
Fragment: 78-87
Site: 14-3-3_pThr83
Modification: (TPO)83
Sequence: ANVPQTVPMR
Construct: ANVPQ(pThr)VPMR (crude synthetic peptide)

Protein B

Protein: YWHAQ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3tau
Average holdup BI: 0.7
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 130
P value: 6.96
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.26
DOI: 10.1101/2022.12.08.519103
Experimental Details

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