Interaction Details


Details of experiments between O96013 and P62258
Note that the results of all experiments are listed, regardless of the modification states of the fragments.


Experiment series 1

Protein A

Protein: PAK4
Fragment: 162-171
Site: 14-3-3_pSer167
Modification: (SEP)167
Sequence: SSREGSGGPQ
Construct: SSREG(pSer)GGPQ (crude synthetic peptide)

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: -0.26
Holdup BI standard deviation: 0.89
Immobilized partner concentration (10-6M): 143
P value: 0.2
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 2

Protein A

Protein: PAK4
Fragment: 176-185
Site: 14-3-3_pSer181
Modification: (SEP)181
Sequence: DKRPLSGPDV
Construct: DKRPL(pSer)GPDV (crude synthetic peptide)

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.04
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 116
P value: 0.78
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 3

Protein A

Protein: PAK4
Fragment: 176-185
Site: 14-3-3_pSer181
Modification: (SEP)181
Sequence: DKRPLSGPDV
Construct: DKRPL(pSer)GPDV (crude synthetic peptide)

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.04
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 143
P value: 1.35
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 4

Protein A

Protein: PAK4
Fragment: 262-271
Site: 14-3-3_pSer267
Modification: (SEP)267
Sequence: LGPHASEPQL
Construct: LGPHA(pSer)EPQL (crude synthetic peptide)

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.35
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 116
P value: 7.54
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 5

Protein A

Protein: PAK4
Fragment: 262-271
Site: 14-3-3_pSer267
Modification: (SEP)267
Sequence: LGPHASEPQL
Construct: LGPHA(pSer)EPQL (crude synthetic peptide)

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.37
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 143
P value: 5.44
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.62
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: PAK4
Fragment: 36-45
Site: 14-3-3_pSer41
Modification: (SEP)41
Sequence: PRQWQSLIEE
Construct: PRQWQ(pSer)LIEE (crude synthetic peptide)

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.15
Holdup BI standard deviation: 0.23
Immobilized partner concentration (10-6M): 116
P value: 0.59
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 7

Protein A

Protein: PAK4
Fragment: 469-478
Site: 14-3-3_pSer474
Modification: (SEP)474
Sequence: VPRRKSLVGT
Construct: VPRRK(pSer)LVGT (crude synthetic peptide)

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.1
Holdup BI standard deviation: 0.08
Immobilized partner concentration (10-6M): 116
P value: 1.26
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 8

Protein A

Protein: PAK4
Fragment: 469-478
Site: 14-3-3_pSer474
Modification: (SEP)474
Sequence: VPRRKSLVGT
Construct: VPRRK(pSer)LVGT (crude synthetic peptide)

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.08
Holdup BI standard deviation: 0.1
Immobilized partner concentration (10-6M): 143
P value: 0.92
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: PAK4
Fragment: 202-211
Site: 14-3-3_pThr207
Modification: (TPO)207
Sequence: GRPFNTYPRA
Construct: GRPFN(pThr)YPRA (crude synthetic peptide)

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.83
Holdup BI standard deviation: 0.17
Immobilized partner concentration (10-6M): 116
P value: 2.37
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.64
DOI: 10.1101/2022.12.08.519103
Experimental Details

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