Interaction Details


Details of experiments between O95208 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: EPN2
Fragment: 168-177
Site: 14-3-3_pSer173
Modification: (SEP)173
Sequence: FGRGSSQPNL
Construct: FGRGS(pSer)QPNL (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: 0.52
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 143
P value: 6.99
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.88
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: EPN2
Fragment: 168-177
Site: 14-3-3_pSer173
Modification: (SEP)173
Sequence: FGRGSSQPNL
Construct: FGRGS(pSer)QPNL (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: 0.46
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 116
P value: 5.93
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.87
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: EPN2
Fragment: 190-199
Site: 14-3-3_pSer195
Modification: (SEP)195
Sequence: GGSPASYHGS
Construct: GGSPA(pSer)YHGS (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: 0.41
Holdup BI standard deviation: 0.1
Immobilized partner concentration (10-6M): 143
P value: 4.04
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.69
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: EPN2
Fragment: 190-199
Site: 14-3-3_pSer195
Modification: (SEP)195
Sequence: GGSPASYHGS
Construct: GGSPA(pSer)YHGS (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: 0.19
Holdup BI standard deviation: 0.11
Immobilized partner concentration (10-6M): 116
P value: 1.51
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 5

Protein A

Protein: EPN2
Fragment: 322-331
Site: 14-3-3_pSer327
Modification: (SEP)327
Sequence: KKEHGSLPQQ
Construct: KKEHG(pSer)LPQQ (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: -0.01
Holdup BI standard deviation: 0.33
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: EPN2
Fragment: 322-331
Site: 14-3-3_pSer327
Modification: (SEP)327
Sequence: KKEHGSLPQQ
Construct: KKEHG(pSer)LPQQ (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: 0.43
Immobilized partner concentration (10-6M): 143
P value: 0.23
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: EPN2
Fragment: 403-412
Site: 14-3-3_pSer408
Modification: (SEP)408
Sequence: SVPKNSDPWA
Construct: SVPKN(pSer)DPWA (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: 0.18
Holdup BI standard deviation: 0.32
Immobilized partner concentration (10-6M): 116
P value: 0.53
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: EPN2
Fragment: 403-412
Site: 14-3-3_pSer408
Modification: (SEP)408
Sequence: SVPKNSDPWA
Construct: SVPKN(pSer)DPWA (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: 0.02
Holdup BI standard deviation: 0.08
Immobilized partner concentration (10-6M): 143
P value: 0.15
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: EPN2
Fragment: 267-276
Site: 14-3-3_pThr272
Modification: (TPO)272
Sequence: QARPQTSGEE
Construct: QARPQ(pThr)SGEE (crude synthetic peptide)

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Measured dissociation constant (10-6M): <100
Immobilized partner concentration (10-6M): 116
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 1
Measured pKd: 4
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: EPN2
Fragment: 311-320
Site: 14-3-3_pThr316
Modification: (TPO)316
Sequence: ESRRDTVKIP
Construct: ESRRD(pThr)VKIP (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
Holdup BI standard deviation: 0.11
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 11

Protein A

Protein: EPN2
Fragment: 311-320
Site: 14-3-3_pThr316
Modification: (TPO)316
Sequence: ESRRDTVKIP
Construct: ESRRD(pThr)VKIP (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: -0.06
Holdup BI standard deviation: 0.22
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))