Interaction Details


Details of experiments between P31947 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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: -0.13
Holdup BI standard deviation: 0.9
Immobilized partner concentration (10-6M): 142
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: 0.02
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 113
P value: 0.29
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: 0.05
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 142
P value: 2.72
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: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: 0.04
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 111
P value: 0.76
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 5

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: 0.19
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 113
P value: 5.6
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 6

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: 0.2
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 142
P value: 3.32
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: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: 0.14
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 111
P value: 3.31
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: PAK4
Fragment: 36-45
Site: 14-3-3_pSer41
Modification: (SEP)41
Sequence: PRQWQSLIEE
Construct: PRQWQ(pSer)LIEE (crude synthetic peptide)
Average holdup BI: 0.05
Holdup BI standard deviation: 0.27
Immobilized partner concentration (10-6M): 113
P value: 0.15
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: PAK4
Fragment: 36-45
Site: 14-3-3_pSer41
Modification: (SEP)41
Sequence: PRQWQSLIEE
Construct: PRQWQ(pSer)LIEE (crude synthetic peptide)
Average holdup BI: -0.15
Holdup BI standard deviation: 0.43
Immobilized partner concentration (10-6M): 142
P value: 0.39
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: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: PAK4
Fragment: 36-45
Site: 14-3-3_pSer41
Modification: (SEP)41
Sequence: PRQWQSLIEE
Construct: PRQWQ(pSer)LIEE (crude synthetic peptide)
Average holdup BI: 0.19
Holdup BI standard deviation: 0.11
Immobilized partner concentration (10-6M): 111
P value: 1.69
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: 0.02
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 113
P value: 0.31
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 12

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: 0.1
Immobilized partner concentration (10-6M): 142
P value: 0.7
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: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

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
Holdup BI standard deviation: 0.18
Immobilized partner concentration (10-6M): 111
P value: 0.02
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 14

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: PAK4
Fragment: 483-492
Site: 14-3-3_pSer488
Modification: (SEP)488
Sequence: APELISRLPY
Construct: APELI(pSer)RLPY (crude synthetic peptide)
Average holdup BI: -0.21
Holdup BI standard deviation: 0.31
Immobilized partner concentration (10-6M): 113
P value: 0.67
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 15

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: PAK4
Fragment: 483-492
Site: 14-3-3_pSer488
Modification: (SEP)488
Sequence: APELISRLPY
Construct: APELI(pSer)RLPY (crude synthetic peptide)
Average holdup BI: -0.11
Holdup BI standard deviation: 0.09
Immobilized partner concentration (10-6M): 111
P value: 1.41
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 16

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

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): 113
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
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 4))

Experiment series 17

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

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): 142
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))

Experiment series 18

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: PAK4
Fragment: 202-211
Site: 14-3-3_pThr207
Modification: (TPO)207
Sequence: GRPFNTYPRA
Construct: GRPFN(pThr)YPRA (crude synthetic peptide)
Average holdup BI: 0.93
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 111
P value: 2.55
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 5.08
DOI: 10.1101/2022.12.08.519103
Experimental Details

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