Interaction Details


Details of experiments between Q12778 and P61981
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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Measured dissociation constant (10-6M):
Standard deviation of dissociation constant (10-6M): 0
Average holdup BI: 0.46
Holdup BI standard deviation: 0.06
Normalized holdup BI: 0
Normalized holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 129
P value: 5.56
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.82
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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Measured dissociation constant (10-6M):
Standard deviation of dissociation constant (10-6M): 0
Average holdup BI: -0.08
Holdup BI standard deviation: 0.37
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 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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Measured dissociation constant (10-6M):
Standard deviation of dissociation constant (10-6M): 0
Average holdup BI: -0.15
Holdup BI standard deviation: 0.45
Normalized holdup BI: 0
Normalized holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 134
P value: 0.29
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: 271-280
Site: 14-3-3_pSer276
Modification: (SEP)276
Sequence: AKKKASLQSG
Construct: AKKKA(pSer)LQSG (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Measured dissociation constant (10-6M):
Standard deviation of dissociation constant (10-6M): 0
Average holdup BI: 0.56
Holdup BI standard deviation: 0.16
Normalized holdup BI: 0
Normalized holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 129
P value: 3.63
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.99
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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Measured dissociation constant (10-6M):
Standard deviation of dissociation constant (10-6M): 0
Average holdup BI: -0.14
Holdup BI standard deviation: 1.02
Normalized holdup BI: 0
Normalized holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 134
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 6

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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Measured dissociation constant (10-6M):
Standard deviation of dissociation constant (10-6M): 0
Average holdup BI: -0.14
Holdup BI standard deviation: 1.01
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 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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Measured dissociation constant (10-6M):
Standard deviation of dissociation constant (10-6M): 0
Average holdup BI: -0.03
Holdup BI standard deviation: 0.61
Normalized holdup BI: 0
Normalized holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 129
P value: 0.02
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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Measured dissociation constant (10-6M):
Standard deviation of dissociation constant (10-6M): 0
Average holdup BI: 0.06
Holdup BI standard deviation: 0.04
Normalized holdup BI: 0
Normalized holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 134
P value: 1.71
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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Measured dissociation constant (10-6M):
Standard deviation of dissociation constant (10-6M): 0
Average holdup BI: 0.03
Holdup BI standard deviation: 0.02
Normalized holdup BI: 0
Normalized holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 129
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
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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Measured dissociation constant (10-6M):
Standard deviation of dissociation constant (10-6M): 0
Average holdup BI: -0.12
Holdup BI standard deviation: 0.05
Normalized holdup BI: 0
Normalized holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 134
P value: 2.65
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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Measured dissociation constant (10-6M):
Standard deviation of dissociation constant (10-6M): 0
Average holdup BI: 0.07
Holdup BI standard deviation: 0.09
Normalized holdup BI: 0
Normalized holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 129
P value: 0.84
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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Measured dissociation constant (10-6M):
Standard deviation of dissociation constant (10-6M): 0
Average holdup BI: 0.1
Holdup BI standard deviation: 0.47
Normalized holdup BI: 0
Normalized holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 134
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 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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Measured dissociation constant (10-6M):
Standard deviation of dissociation constant (10-6M): 0
Average holdup BI: 0.16
Holdup BI standard deviation: 0.07
Normalized holdup BI: 0
Normalized holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 129
P value: 2.53
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: FOXO1
Fragment: 213-222
Site: 14-3-3_pSer218
Modification: (SEP)218
Sequence: IRHNLSLHSK
Construct: IRHNL(pSer)LHSK (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Measured dissociation constant (10-6M):
Standard deviation of dissociation constant (10-6M): 0
Average holdup BI: 0.59
Holdup BI standard deviation: 0.26
Normalized holdup BI: 0
Normalized holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 115
P value: 1.46
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.11
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: FOXO1
Fragment: 271-280
Site: 14-3-3_pSer276
Modification: (SEP)276
Sequence: AKKKASLQSG
Construct: AKKKA(pSer)LQSG (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Measured dissociation constant (10-6M):
Standard deviation of dissociation constant (10-6M): 0
Average holdup BI: 0.75
Holdup BI standard deviation: 0.23
Normalized holdup BI: 0
Normalized holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 115
P value: 2.69
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.44
DOI: 10.1101/2022.12.08.519103
Experimental Details

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


Experiment series 16

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: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Measured dissociation constant (10-6M):
Standard deviation of dissociation constant (10-6M): 0
Average holdup BI: -0.15
Holdup BI standard deviation: 0.62
Normalized holdup BI: 0
Normalized holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 115
P value: 0.19
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: FOXO1
Fragment: 320-329
Site: 14-3-3_pSer325
Modification: (SEP)325
Sequence: NASTISGRLS
Construct: NASTI(pSer)GRLS (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Measured dissociation constant (10-6M):
Standard deviation of dissociation constant (10-6M): 0
Average holdup BI: 0.05
Holdup BI standard deviation: 0.04
Normalized holdup BI: 0
Normalized holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 115
P value: 1.12
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 18

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: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Measured dissociation constant (10-6M):
Standard deviation of dissociation constant (10-6M): 0
Average holdup BI: 0.2
Holdup BI standard deviation: 0.1
Normalized holdup BI: 0
Normalized holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 115
P value: 1.81
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 19

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: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Measured dissociation constant (10-6M):
Standard deviation of dissociation constant (10-6M): 0
Average holdup BI: -0.05
Holdup BI standard deviation: 1.31
Normalized holdup BI: 0
Normalized holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 115
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))