Interaction Details


Details of experiments between Q12778 and P31946
Note that the results of all experiments are listed, regardless of the modification states of the fragments.


Experiment series 1

Protein A

Protein: FOXO1
Fragment: 213-222
Site: 14-3-3_pSer218
Modification: (SEP)218
Sequence: IRHNLSLHSK
Construct: IRHNL(pSer)LHSK (crude synthetic peptide)

Protein B

Protein: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Measured dissociation constant (10-6M):
Standard deviation of dissociation constant (10-6M): 0
Average holdup BI: 0.37
Holdup BI standard deviation: 0.25
Normalized holdup BI: 0
Normalized holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 102
P value: 1.64
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.76
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: FOXO1
Fragment: 213-222
Site: 14-3-3_pSer218
Modification: (SEP)218
Sequence: IRHNLSLHSK
Construct: IRHNL(pSer)LHSK (crude synthetic peptide)

Protein B

Protein: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Measured dissociation constant (10-6M):
Standard deviation of dissociation constant (10-6M): 0
Average holdup BI: -0.45
Holdup BI standard deviation: 1.32
Normalized holdup BI: 0
Normalized holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 134
P value: 0.3
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 3

Protein A

Protein: FOXO1
Fragment: 251-260
Site: 14-3-3_pSer256
Modification: (SEP)256
Sequence: RRRAASMDNN
Construct: RRRAA(pSer)MDNN (crude synthetic peptide)

Protein B

Protein: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Measured dissociation constant (10-6M):
Standard deviation of dissociation constant (10-6M): 0
Average holdup BI: -0.08
Holdup BI standard deviation: 0.31
Normalized holdup BI: 0
Normalized holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 134
P value: 0.23
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: FOXO1
Fragment: 251-260
Site: 14-3-3_pSer256
Modification: (SEP)256
Sequence: RRRAASMDNN
Construct: RRRAA(pSer)MDNN (crude synthetic peptide)

Protein B

Protein: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Measured dissociation constant (10-6M):
Standard deviation of dissociation constant (10-6M): 0
Average holdup BI: -0.04
Holdup BI standard deviation: 0.86
Normalized holdup BI: 0
Normalized holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 102
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 5

Protein A

Protein: FOXO1
Fragment: 271-280
Site: 14-3-3_pSer276
Modification: (SEP)276
Sequence: AKKKASLQSG
Construct: AKKKA(pSer)LQSG (crude synthetic peptide)

Protein B

Protein: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Measured dissociation constant (10-6M):
Standard deviation of dissociation constant (10-6M): 0
Average holdup BI: 0.66
Holdup BI standard deviation: 0.42
Normalized holdup BI: 0
Normalized holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 102
P value: 1.65
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.27
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: FOXO1
Fragment: 271-280
Site: 14-3-3_pSer276
Modification: (SEP)276
Sequence: AKKKASLQSG
Construct: AKKKA(pSer)LQSG (crude synthetic peptide)

Protein B

Protein: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Measured dissociation constant (10-6M):
Standard deviation of dissociation constant (10-6M): 0
Average holdup BI: -0.27
Holdup BI standard deviation: 1.28
Normalized holdup BI: 0
Normalized holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 134
P value: 0.2
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: FOXO1
Fragment: 317-326
Site: 14-3-3_pSer322
Modification: (SEP)322
Sequence: TSSNASTISG
Construct: TSSNA(pSer)TISG (crude synthetic peptide)

Protein B

Protein: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Measured dissociation constant (10-6M):
Standard deviation of dissociation constant (10-6M): 0
Average holdup BI: 0.02
Holdup BI standard deviation: 0.33
Normalized holdup BI: 0
Normalized holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 102
P value: 0.03
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: FOXO1
Fragment: 320-329
Site: 14-3-3_pSer325
Modification: (SEP)325
Sequence: NASTISGRLS
Construct: NASTI(pSer)GRLS (crude synthetic peptide)

Protein B

Protein: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Measured dissociation constant (10-6M):
Standard deviation of dissociation constant (10-6M): 0
Average holdup BI: 0
Holdup BI standard deviation: 0.04
Normalized holdup BI: 0
Normalized holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 134
P value: 0.08
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: FOXO1
Fragment: 320-329
Site: 14-3-3_pSer325
Modification: (SEP)325
Sequence: NASTISGRLS
Construct: NASTI(pSer)GRLS (crude synthetic peptide)

Protein B

Protein: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Measured dissociation constant (10-6M):
Standard deviation of dissociation constant (10-6M): 0
Average holdup BI: 0.02
Holdup BI standard deviation: 0.04
Normalized holdup BI: 0
Normalized holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 102
P value: 0.64
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: FOXO1
Fragment: 177-186
Site: 14-3-3_pThr182
Modification: (TPO)182
Sequence: AEKRLTLSQI
Construct: AEKRL(pThr)LSQI (crude synthetic peptide)

Protein B

Protein: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Measured dissociation constant (10-6M):
Standard deviation of dissociation constant (10-6M): 0
Average holdup BI: 0.19
Holdup BI standard deviation: 0.11
Normalized holdup BI: 0
Normalized holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 134
P value: 1.86
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: FOXO1
Fragment: 177-186
Site: 14-3-3_pThr182
Modification: (TPO)182
Sequence: AEKRLTLSQI
Construct: AEKRL(pThr)LSQI (crude synthetic peptide)

Protein B

Protein: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Measured dissociation constant (10-6M):
Standard deviation of dissociation constant (10-6M): 0
Average holdup BI: 0.06
Holdup BI standard deviation: 0.09
Normalized holdup BI: 0
Normalized holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 102
P value: 0.68
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 12

Protein A

Protein: FOXO1
Fragment: 644-653
Site: 14-3-3_pThr649
Modification: (TPO)649
Sequence: SVKTTTHSWV
Construct: SVKTT(pThr)HSWV (crude synthetic peptide)

Protein B

Protein: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Measured dissociation constant (10-6M):
Standard deviation of dissociation constant (10-6M): 0
Average holdup BI: 0.1
Holdup BI standard deviation: 0.56
Normalized holdup BI: 0
Normalized holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 134
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 13

Protein A

Protein: FOXO1
Fragment: 644-653
Site: 14-3-3_pThr649
Modification: (TPO)649
Sequence: SVKTTTHSWV
Construct: SVKTT(pThr)HSWV (crude synthetic peptide)

Protein B

Protein: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Measured dissociation constant (10-6M):
Standard deviation of dissociation constant (10-6M): 0
Average holdup BI: 0.08
Holdup BI standard deviation: 0.11
Normalized holdup BI: 0
Normalized holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 102
P value: 0.81
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))