Interaction Details


Details of experiments between P31947 and Q99759
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: MAP3K3
Fragment: 142-151
Site: 14-3-3_pSer147
Modification: (SEP)147
Sequence: IKASQSAGDI
Construct: IKASQ(pSer)AGDI (crude synthetic peptide)
Average holdup BI: 0.02
Holdup BI standard deviation: 0.09
Immobilized partner concentration (10-6M): 113
P value: 0.13
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 2

Protein A

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

Protein B

Protein: MAP3K3
Fragment: 142-151
Site: 14-3-3_pSer147
Modification: (SEP)147
Sequence: IKASQSAGDI
Construct: IKASQ(pSer)AGDI (crude synthetic peptide)
Average holdup BI: 0.04
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 142
P value: 0.74
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: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: MAP3K3
Fragment: 142-151
Site: 14-3-3_pSer147
Modification: (SEP)147
Sequence: IKASQSAGDI
Construct: IKASQ(pSer)AGDI (crude synthetic peptide)
Average holdup BI: -0.02
Holdup BI standard deviation: 0.11
Immobilized partner concentration (10-6M): 111
P value: 0.16
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 4

Protein A

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

Protein B

Protein: MAP3K3
Fragment: 161-170
Site: 14-3-3_pSer166
Modification: (SEP)166
Sequence: RSRHLSVSSQ
Construct: RSRHL(pSer)VSSQ (crude synthetic peptide)
Average holdup BI: 0.96
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 113
P value: 3.74
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 5.28
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: MAP3K3
Fragment: 245-254
Site: 14-3-3_pSer250
Modification: (SEP)250
Sequence: MSRAQSFPDN
Construct: MSRAQ(pSer)FPDN (crude synthetic peptide)
Average holdup BI: 0.65
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 142
P value: 10.01
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.12
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: MAP3K3
Fragment: 245-254
Site: 14-3-3_pSer250
Modification: (SEP)250
Sequence: MSRAQSFPDN
Construct: MSRAQ(pSer)FPDN (crude synthetic peptide)
Average holdup BI: 0.58
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 113
P value: 4.79
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.08
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: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: MAP3K3
Fragment: 245-254
Site: 14-3-3_pSer250
Modification: (SEP)250
Sequence: MSRAQSFPDN
Construct: MSRAQ(pSer)FPDN (crude synthetic peptide)
Average holdup BI: 0.44
Holdup BI standard deviation: 0.12
Immobilized partner concentration (10-6M): 111
P value: 3.33
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.85
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: MAP3K3
Fragment: 332-341
Site: 14-3-3_pSer337
Modification: (SEP)337
Sequence: RGRLRSADSE
Construct: RGRLR(pSer)ADSE (crude synthetic peptide)
Average holdup BI: 0.21
Holdup BI standard deviation: 0.33
Immobilized partner concentration (10-6M): 142
P value: 0.52
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: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: MAP3K3
Fragment: 332-341
Site: 14-3-3_pSer337
Modification: (SEP)337
Sequence: RGRLRSADSE
Construct: RGRLR(pSer)ADSE (crude synthetic peptide)
Average holdup BI: -0.14
Holdup BI standard deviation: 1.54
Immobilized partner concentration (10-6M): 111
P value: 0.06
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 10

Protein A

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

Protein B

Protein: MAP3K3
Fragment: 335-344
Site: 14-3-3_pSer340
Modification: (SEP)340
Sequence: LRSADSENAL
Construct: LRSAD(pSer)ENAL (crude synthetic peptide)
Average holdup BI: 0.03
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 113
P value: 0.7
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: MAP3K3
Fragment: 335-344
Site: 14-3-3_pSer340
Modification: (SEP)340
Sequence: LRSADSENAL
Construct: LRSAD(pSer)ENAL (crude synthetic peptide)
Average holdup BI: -0.02
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 142
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: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: MAP3K3
Fragment: 335-344
Site: 14-3-3_pSer340
Modification: (SEP)340
Sequence: LRSADSENAL
Construct: LRSAD(pSer)ENAL (crude synthetic peptide)
Average holdup BI: 0
Holdup BI standard deviation: 0.03
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 13

Protein A

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

Protein B

Protein: MAP3K3
Fragment: 352-361
Site: 14-3-3_pSer357
Modification: (SEP)357
Sequence: PTKSPSAPIN
Construct: PTKSP(pSer)APIN (crude synthetic peptide)
Average holdup BI: 0.47
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 113
P value: 6.83
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.9
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: MAP3K3
Fragment: 352-361
Site: 14-3-3_pSer357
Modification: (SEP)357
Sequence: PTKSPSAPIN
Construct: PTKSP(pSer)APIN (crude synthetic peptide)
Average holdup BI: 0.51
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 142
P value: 6.28
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.86
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

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: MAP3K3
Fragment: 352-361
Site: 14-3-3_pSer357
Modification: (SEP)357
Sequence: PTKSPSAPIN
Construct: PTKSP(pSer)APIN (crude synthetic peptide)
Average holdup BI: 0.41
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 111
P value: 5.67
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.79
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: MAP3K3
Fragment: 289-298
Site: 14-3-3_pThr294
Modification: (TPO)294
Sequence: SDGRRTFPRI
Construct: SDGRR(pThr)FPRI (crude synthetic peptide)
Average holdup BI: 0.75
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 142
P value: 6.07
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.32
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

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: MAP3K3
Fragment: 289-298
Site: 14-3-3_pThr294
Modification: (TPO)294
Sequence: SDGRRTFPRI
Construct: SDGRR(pThr)FPRI (crude synthetic peptide)
Average holdup BI: 0.67
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 113
P value: 5.88
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.24
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

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: MAP3K3
Fragment: 289-298
Site: 14-3-3_pThr294
Modification: (TPO)294
Sequence: SDGRRTFPRI
Construct: SDGRR(pThr)FPRI (crude synthetic peptide)
Average holdup BI: 0.69
Holdup BI standard deviation: 0.09
Immobilized partner concentration (10-6M): 111
P value: 3.85
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.29
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 19

Protein A

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

Protein B

Protein: MAP3K3
Fragment: 523-532
Site: 14-3-3_pThr528
Modification: (TPO)528
Sequence: GMRSVTGTPY
Construct: GMRSV(pThr)GTPY (crude synthetic peptide)
Average holdup BI: 0.4
Holdup BI standard deviation: 0.11
Immobilized partner concentration (10-6M): 113
P value: 3.03
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.77
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 20

Protein A

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

Protein B

Protein: MAP3K3
Fragment: 523-532
Site: 14-3-3_pThr528
Modification: (TPO)528
Sequence: GMRSVTGTPY
Construct: GMRSV(pThr)GTPY (crude synthetic peptide)
Average holdup BI: 0.37
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 142
P value: 4.55
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))