Interaction Details


Details of experiments between O96013 and P31946
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: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.06
Holdup BI standard deviation: 0.58
Immobilized partner concentration (10-6M): 102
P value: 0.06
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: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.14
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 134
P value: 2.96
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: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.12
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 102
P value: 2.76
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: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.5
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 102
P value: 6.09
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4
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: PAK4
Fragment: 262-271
Site: 14-3-3_pSer267
Modification: (SEP)267
Sequence: LGPHASEPQL
Construct: LGPHA(pSer)EPQL (crude synthetic peptide)

Protein B

Protein: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.52
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 134
P value: 8.39
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.92
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: PAK4
Fragment: 36-45
Site: 14-3-3_pSer41
Modification: (SEP)41
Sequence: PRQWQSLIEE
Construct: PRQWQ(pSer)LIEE (crude synthetic peptide)

Protein B

Protein: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.73
Holdup BI standard deviation: 0.1
Immobilized partner concentration (10-6M): 134
P value: 4.19
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.3
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: 36-45
Site: 14-3-3_pSer41
Modification: (SEP)41
Sequence: PRQWQSLIEE
Construct: PRQWQ(pSer)LIEE (crude synthetic peptide)

Protein B

Protein: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.65
Holdup BI standard deviation: 0.17
Immobilized partner concentration (10-6M): 102
P value: 2.28
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.25
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: PAK4
Fragment: 469-478
Site: 14-3-3_pSer474
Modification: (SEP)474
Sequence: VPRRKSLVGT
Construct: VPRRK(pSer)LVGT (crude synthetic peptide)

Protein B

Protein: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.18
Holdup BI standard deviation: 0.08
Immobilized partner concentration (10-6M): 134
P value: 2.19
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 9

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: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.15
Holdup BI standard deviation: 0.11
Immobilized partner concentration (10-6M): 102
P value: 1.45
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: PAK4
Fragment: 483-492
Site: 14-3-3_pSer488
Modification: (SEP)488
Sequence: APELISRLPY
Construct: APELI(pSer)RLPY (crude synthetic peptide)

Protein B

Protein: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.03
Holdup BI standard deviation: 0.23
Immobilized partner concentration (10-6M): 134
P value: 0.11
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 11

Protein A

Protein: PAK4
Fragment: 483-492
Site: 14-3-3_pSer488
Modification: (SEP)488
Sequence: APELISRLPY
Construct: APELI(pSer)RLPY (crude synthetic peptide)

Protein B

Protein: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: -0.11
Holdup BI standard deviation: 0.48
Immobilized partner concentration (10-6M): 102
P value: 0.21
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: PAK4
Fragment: 202-211
Site: 14-3-3_pThr207
Modification: (TPO)207
Sequence: GRPFNTYPRA
Construct: GRPFN(pThr)YPRA (crude synthetic peptide)

Protein B

Protein: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.69
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 134
P value: 2.35
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.23
DOI: 10.1101/2022.12.08.519103
Experimental Details

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