Interaction Details


Details of experiments between P31946 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: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta

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: 0.07
Immobilized partner concentration (10-6M): 134
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 2

Protein A

Protein: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta

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: 0.1
Holdup BI standard deviation: 0.14
Immobilized partner concentration (10-6M): 102
P value: 0.67
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: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta

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: 0.94
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 134
P value: 3.73
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 5.07
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: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta

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: 0.72
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 102
P value: 13.37
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.39
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: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta

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: 0.71
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 134
P value: 5.75
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.27
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: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta

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: -0.26
Holdup BI standard deviation: 1.09
Immobilized partner concentration (10-6M): 102
P value: 0.16
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: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta

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: 0.04
Immobilized partner concentration (10-6M): 134
P value: 0.35
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: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta

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: 0.01
Holdup BI standard deviation: 0.08
Immobilized partner concentration (10-6M): 102
P value: 0.07
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: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta

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: 0.74
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 102
P value: 7.74
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.45
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: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta

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: 0.76
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 134
P value: 7.66
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.37
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: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta

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: 0.87
Holdup BI standard deviation: 0.13
Immobilized partner concentration (10-6M): 134
P value: 4.77
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.68
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: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta

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: 0.79
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 102
P value: 6.54
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.56
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: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta

Protein B

Protein: MAP3K3
Fragment: 511-520
Site: 14-3-3_pThr516
Modification: (TPO)516
Sequence: SKRLQTICMS
Construct: SKRLQ(pThr)ICMS (crude synthetic peptide)
Average holdup BI: 0.01
Holdup BI standard deviation: 0.76
Immobilized partner concentration (10-6M): 134
P value: 0.01
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: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta

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: 0.71
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 134
P value: 5.18
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.26
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: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta

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: 0.61
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 102
P value: 6.06
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.19
DOI: 10.1101/2022.12.08.519103
Experimental Details

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