Interaction Details


Details of experiments between P62258 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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

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): 116
P value: 0.43
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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

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
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 143
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 3

Protein A

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

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: 0.88
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 116
P value: 3.53
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.8
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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

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: 0.54
Holdup BI standard deviation: 0.08
Immobilized partner concentration (10-6M): 116
P value: 4.39
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.01
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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

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: 0.59
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 143
P value: 10.47
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 6

Protein A

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

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: 0.11
Holdup BI standard deviation: 0.12
Immobilized partner concentration (10-6M): 116
P value: 0.94
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 7

Protein A

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

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): 143
P value: 0.2
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 8

Protein A

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

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: 0.54
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 116
P value: 7.49
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.01
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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

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: 0.57
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 143
P value: 6.48
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.96
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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

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: 0.03
Immobilized partner concentration (10-6M): 143
P value: 6.68
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.33
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 11

Protein A

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

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: 0.66
Holdup BI standard deviation: 0.08
Immobilized partner concentration (10-6M): 116
P value: 5.04
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))

Experiment series 12

Protein A

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

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: 0.57
Holdup BI standard deviation: 0.1
Immobilized partner concentration (10-6M): 116
P value: 3.88
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.05
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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

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: 0.57
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 143
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: 3.97
DOI: 10.1101/2022.12.08.519103
Experimental Details

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