Interaction Details


Details of experiments between Q9Y2U5 and P62258
Note that the results of all experiments are listed, regardless of the modification states of the fragments.


Experiment series 1

Protein A

Protein: MAP3K2
Fragment: 148-157
Site: 14-3-3_pSer153
Modification: (SEP)153
Sequence: RKKRLSIIGP
Construct: RKKRL(pSer)IIGP (crude synthetic peptide)

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: -0.31
Holdup BI standard deviation: 0.45
Immobilized partner concentration (10-6M): 116
P value: 0.68
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: MAP3K2
Fragment: 234-243
Site: 14-3-3_pSer239
Modification: (SEP)239
Sequence: MPRAQSYPDN
Construct: MPRAQ(pSer)YPDN (crude synthetic peptide)

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.69
Holdup BI standard deviation: 0.09
Immobilized partner concentration (10-6M): 116
P value: 4.67
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 3

Protein A

Protein: MAP3K2
Fragment: 234-243
Site: 14-3-3_pSer239
Modification: (SEP)239
Sequence: MPRAQSYPDN
Construct: MPRAQ(pSer)YPDN (crude synthetic peptide)

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.65
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 143
P value: 9.6
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 4

Protein A

Protein: MAP3K2
Fragment: 243-252
Site: 14-3-3_pSer248
Modification: (SEP)248
Sequence: NHQEFSDYDN
Construct: NHQEF(pSer)DYDN (crude synthetic peptide)

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.12
Holdup BI standard deviation: 0.48
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 5

Protein A

Protein: MAP3K2
Fragment: 326-335
Site: 14-3-3_pSer331
Modification: (SEP)331
Sequence: IRRRGSDIDN
Construct: IRRRG(pSer)DIDN (crude synthetic peptide)

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: -0.1
Holdup BI standard deviation: 1.49
Immobilized partner concentration (10-6M): 116
P value: 0.03
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: MAP3K2
Fragment: 488-497
Site: 14-3-3_pSer493
Modification: (SEP)493
Sequence: NILRDSTGNV
Construct: NILRD(pSer)TGNV (crude synthetic peptide)

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: -0.04
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 116
P value: 2.2
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: MAP3K2
Fragment: 488-497
Site: 14-3-3_pSer493
Modification: (SEP)493
Sequence: NILRDSTGNV
Construct: NILRD(pSer)TGNV (crude synthetic peptide)

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.03
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 143
P value: 0.86
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: MAP3K2
Fragment: 515-524
Site: 14-3-3_pSer520
Modification: (SEP)520
Sequence: GTGMKSVTGT
Construct: GTGMK(pSer)VTGT (crude synthetic peptide)

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.11
Holdup BI standard deviation: 0.25
Immobilized partner concentration (10-6M): 116
P value: 0.4
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: MAP3K2
Fragment: 515-524
Site: 14-3-3_pSer520
Modification: (SEP)520
Sequence: GTGMKSVTGT
Construct: GTGMK(pSer)VTGT (crude synthetic peptide)

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.04
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 143
P value: 0.83
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: MAP3K2
Fragment: 258-267
Site: 14-3-3_pThr263
Modification: (TPO)263
Sequence: FGKGGTYPRR
Construct: FGKGG(pThr)YPRR (crude synthetic peptide)

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.8
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 143
P value: 6.84
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 11

Protein A

Protein: MAP3K2
Fragment: 258-267
Site: 14-3-3_pThr263
Modification: (TPO)263
Sequence: FGKGGTYPRR
Construct: FGKGG(pThr)YPRR (crude synthetic peptide)

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.73
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 116
P value: 5.05
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 12

Protein A

Protein: MAP3K2
Fragment: 278-287
Site: 14-3-3_pThr283
Modification: (TPO)283
Sequence: NDGRKTFPRA
Construct: NDGRK(pThr)FPRA (crude synthetic peptide)

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.65
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 143
P value: 6.57
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.11
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: MAP3K2
Fragment: 278-287
Site: 14-3-3_pThr283
Modification: (TPO)283
Sequence: NDGRKTFPRA
Construct: NDGRK(pThr)FPRA (crude synthetic peptide)

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.57
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 116
P value: 4.71
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.06
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: MAP3K2
Fragment: 517-526
Site: 14-3-3_pThr522
Modification: (TPO)522
Sequence: GMKSVTGTPY
Construct: GMKSV(pThr)GTPY (crude synthetic peptide)

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.54
Holdup BI standard deviation: 0.08
Immobilized partner concentration (10-6M): 143
P value: 4.93
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.91
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: MAP3K2
Fragment: 517-526
Site: 14-3-3_pThr522
Modification: (TPO)522
Sequence: GMKSVTGTPY
Construct: GMKSV(pThr)GTPY (crude synthetic peptide)

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.36
Holdup BI standard deviation: 0.09
Immobilized partner concentration (10-6M): 116
P value: 3.22
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.69
DOI: 10.1101/2022.12.08.519103
Experimental Details

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