Interaction Details


Details of experiments between Q99759 and P27348
Note that the results of all experiments are listed, regardless of the modification states of the fragments.


Experiment series 1

Protein A

Protein: MAP3K3
Fragment: 142-151
Site: 14-3-3_pSer147
Modification: (SEP)147
Sequence: IKASQSAGDI
Construct: IKASQ(pSer)AGDI (crude synthetic peptide)

Protein B

Protein: YWHAQ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3tau
Average holdup BI: 0.03
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 130
P value: 0.55
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: MAP3K3
Fragment: 245-254
Site: 14-3-3_pSer250
Modification: (SEP)250
Sequence: MSRAQSFPDN
Construct: MSRAQ(pSer)FPDN (crude synthetic peptide)

Protein B

Protein: YWHAQ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3tau
Average holdup BI: 0.68
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 130
P value: 11.81
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.22
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: MAP3K3
Fragment: 332-341
Site: 14-3-3_pSer337
Modification: (SEP)337
Sequence: RGRLRSADSE
Construct: RGRLR(pSer)ADSE (crude synthetic peptide)

Protein B

Protein: YWHAQ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3tau
Average holdup BI: 0.14
Holdup BI standard deviation: 0.36
Immobilized partner concentration (10-6M): 130
P value: 0.29
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: MAP3K3
Fragment: 335-344
Site: 14-3-3_pSer340
Modification: (SEP)340
Sequence: LRSADSENAL
Construct: LRSAD(pSer)ENAL (crude synthetic peptide)

Protein B

Protein: YWHAQ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3tau
Average holdup BI: 0.01
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 130
P value: 0.17
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: MAP3K3
Fragment: 352-361
Site: 14-3-3_pSer357
Modification: (SEP)357
Sequence: PTKSPSAPIN
Construct: PTKSP(pSer)APIN (crude synthetic peptide)

Protein B

Protein: YWHAQ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3tau
Average holdup BI: 0.67
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 130
P value: 6.77
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))

Experiment series 6

Protein A

Protein: MAP3K3
Fragment: 289-298
Site: 14-3-3_pThr294
Modification: (TPO)294
Sequence: SDGRRTFPRI
Construct: SDGRR(pThr)FPRI (crude synthetic peptide)

Protein B

Protein: YWHAQ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3tau
Average holdup BI: 0.72
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 130
P value: 6.1
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.3
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: MAP3K3
Fragment: 523-532
Site: 14-3-3_pThr528
Modification: (TPO)528
Sequence: GMRSVTGTPY
Construct: GMRSV(pThr)GTPY (crude synthetic peptide)

Protein B

Protein: YWHAQ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3tau
Average holdup BI: 0.54
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 130
P value: 5.64
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))