Interaction Details


Details of experiments between O43524 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: FOXO3
Fragment: 204-213
Site: 14-3-3_pSer209
Modification: (SEP)209
Sequence: AGWKNSIRHN
Construct: AGWKN(pSer)IRHN (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: 0.23
Immobilized partner concentration (10-6M): 116
P value: 0.02
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: FOXO3
Fragment: 204-213
Site: 14-3-3_pSer209
Modification: (SEP)209
Sequence: AGWKNSIRHN
Construct: AGWKN(pSer)IRHN (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: -0.28
Holdup BI standard deviation: 0.55
Immobilized partner concentration (10-6M): 143
P value: 0.48
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: FOXO3
Fragment: 210-219
Site: 14-3-3_pSer215
Modification: (SEP)215
Sequence: IRHNLSLHSR
Construct: IRHNL(pSer)LHSR (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: 0.14
Immobilized partner concentration (10-6M): 116
P value: 0.41
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 4

Protein A

Protein: FOXO3
Fragment: 210-219
Site: 14-3-3_pSer215
Modification: (SEP)215
Sequence: IRHNLSLHSR
Construct: IRHNL(pSer)LHSR (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: 0.16
Immobilized partner concentration (10-6M): 143
P value: 0.1
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: FOXO3
Fragment: 248-257
Site: 14-3-3_pSer253
Modification: (SEP)253
Sequence: RRRAVSMDNS
Construct: RRRAV(pSer)MDNS (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: -0.21
Holdup BI standard deviation: 1.34
Immobilized partner concentration (10-6M): 116
P value: 0.13
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: FOXO3
Fragment: 275-284
Site: 14-3-3_pSer280
Modification: (SEP)280
Sequence: QTAPESADDS
Construct: QTAPE(pSer)ADDS (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: 0.14
Holdup BI standard deviation: 0.86
Immobilized partner concentration (10-6M): 116
P value: 0.11
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: FOXO3
Fragment: 316-325
Site: 14-3-3_pSer321
Modification: (SEP)321
Sequence: NASTVSGRLS
Construct: NASTV(pSer)GRLS (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: 0.03
Immobilized partner concentration (10-6M): 116
P value: 1.07
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: FOXO3
Fragment: 316-325
Site: 14-3-3_pSer321
Modification: (SEP)321
Sequence: NASTVSGRLS
Construct: NASTV(pSer)GRLS (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: 0.04
Immobilized partner concentration (10-6M): 143
P value: 1.06
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 9

Protein A

Protein: FOXO3
Fragment: 394-403
Site: 14-3-3_pSer399
Modification: (SEP)399
Sequence: ITLPPSQPSP
Construct: ITLPP(pSer)QPSP (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: 1.25
Immobilized partner concentration (10-6M): 116
P value: 0.07
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 10

Protein A

Protein: FOXO3
Fragment: 408-417
Site: 14-3-3_pSer413
Modification: (SEP)413
Sequence: MQRSSSFPYT
Construct: MQRSS(pSer)FPYT (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): 116
P value: 5.27
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.54
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: FOXO3
Fragment: 408-417
Site: 14-3-3_pSer413
Modification: (SEP)413
Sequence: MQRSSSFPYT
Construct: MQRSS(pSer)FPYT (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: 0.78
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 143
P value: 7.42
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.4
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 12

Protein A

Protein: FOXO3
Fragment: 550-559
Site: 14-3-3_pSer555
Modification: (SEP)555
Sequence: LSNSVSNMGL
Construct: LSNSV(pSer)NMGL (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: 0.22
Holdup BI standard deviation: 0.2
Immobilized partner concentration (10-6M): 116
P value: 1.07
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 13

Protein A

Protein: FOXO3
Fragment: 550-559
Site: 14-3-3_pSer555
Modification: (SEP)555
Sequence: LSNSVSNMGL
Construct: LSNSV(pSer)NMGL (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: 0.05
Holdup BI standard deviation: 0.11
Immobilized partner concentration (10-6M): 143
P value: 0.44
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 14

Protein A

Protein: FOXO3
Fragment: 583-592
Site: 14-3-3_pSer588
Modification: (SEP)588
Sequence: LSDSLSGSSL
Construct: LSDSL(pSer)GSSL (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: 0.9
Immobilized partner concentration (10-6M): 116
P value: 0.18
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 15

Protein A

Protein: FOXO3
Fragment: 583-592
Site: 14-3-3_pSer588
Modification: (SEP)588
Sequence: LSDSLSGSSL
Construct: LSDSL(pSer)GSSL (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: 2.23
Immobilized partner concentration (10-6M): 143
P value: 0.04
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 16

Protein A

Protein: FOXO3
Fragment: 174-183
Site: 14-3-3_pThr179
Modification: (TPO)179
Sequence: PDKRLTLSQI
Construct: PDKRL(pThr)LSQI (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: -0.15
Holdup BI standard deviation: 0.17
Immobilized partner concentration (10-6M): 116
P value: 0.96
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 17

Protein A

Protein: FOXO3
Fragment: 174-183
Site: 14-3-3_pThr179
Modification: (TPO)179
Sequence: PDKRLTLSQI
Construct: PDKRL(pThr)LSQI (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: 0.16
Immobilized partner concentration (10-6M): 143
P value: 0.58
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))