Interaction Details


Details of experiments between P27348 and O96013
Note that the results of all experiments are listed, regardless of the modification states of the fragments.


Experiment series 1

Protein A

Protein: YWHAQ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3tau

Protein B

Protein: PAK4
Fragment: 162-171
Site: 14-3-3_pSer167
Modification: (SEP)167
Sequence: SSREGSGGPQ
Construct: SSREG(pSer)GGPQ (crude synthetic peptide)
Average holdup BI: 0.09
Holdup BI standard deviation: 0.65
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 2

Protein A

Protein: YWHAQ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3tau

Protein B

Protein: PAK4
Fragment: 176-185
Site: 14-3-3_pSer181
Modification: (SEP)181
Sequence: DKRPLSGPDV
Construct: DKRPL(pSer)GPDV (crude synthetic peptide)
Average holdup BI: 0.08
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 130
P value: 3.15
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 3

Protein A

Protein: YWHAQ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3tau

Protein B

Protein: PAK4
Fragment: 262-271
Site: 14-3-3_pSer267
Modification: (SEP)267
Sequence: LGPHASEPQL
Construct: LGPHA(pSer)EPQL (crude synthetic peptide)
Average holdup BI: 0.37
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 130
P value: 5.58
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.66
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: YWHAQ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3tau

Protein B

Protein: PAK4
Fragment: 469-478
Site: 14-3-3_pSer474
Modification: (SEP)474
Sequence: VPRRKSLVGT
Construct: VPRRK(pSer)LVGT (crude synthetic peptide)
Average holdup BI: 0.07
Holdup BI standard deviation: 0.11
Immobilized partner concentration (10-6M): 130
P value: 0.65
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: YWHAQ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3tau

Protein B

Protein: PAK4
Fragment: 202-211
Site: 14-3-3_pThr207
Modification: (TPO)207
Sequence: GRPFNTYPRA
Construct: GRPFN(pThr)YPRA (crude synthetic peptide)
Measured dissociation constant (10-6M): <10
Immobilized partner concentration (10-6M): 130
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 1
Measured pKd: 5
DOI: 10.1101/2022.12.08.519103
Experimental Details

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