Interaction Details


Details of experiments between Q13136 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: PPFIA1
Fragment: 1134-1143
Site: 14-3-3_pSer1139
Modification: (SEP)1139
Sequence: FRRAPSWRKK
Construct: FRRAP(pSer)WRKK (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: -0.47
Holdup BI standard deviation: 0.72
Immobilized partner concentration (10-6M): 134
P value: 0.77
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: PPFIA1
Fragment: 1134-1143
Site: 14-3-3_pSer1139
Modification: (SEP)1139
Sequence: FRRAPSWRKK
Construct: FRRAP(pSer)WRKK (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.04
Holdup BI standard deviation: 0.46
Immobilized partner concentration (10-6M): 129
P value: 0.07
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: PPFIA1
Fragment: 525-534
Site: 14-3-3_pSer530
Modification: (SEP)530
Sequence: RPHLGSVPDF
Construct: RPHLG(pSer)VPDF (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.71
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 129
P value: 4.42
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.28
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: PPFIA1
Fragment: 663-672
Site: 14-3-3_pSer668
Modification: (SEP)668
Sequence: RVGSGSLDNL
Construct: biotin-ttds-RVGSG(pSer)LDNL (HPLC-purified 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): 125
Standard deviation of dissociation constant (10-6M): 5
Experiment method: competitive Fluorescence Polarization
Details of affinity fitting: Competitive binding equation in ProFit
Date of measurement: 2022-01-10
Number of measurements: 3
Measured pKd: 3.9
DOI: 10.1101/2022.12.08.519103

Experiment series 5

Protein A

Protein: PPFIA1
Fragment: 663-672
Site: 14-3-3_pSer668
Modification: (SEP)668
Sequence: RVGSGSLDNL
Construct: RVGSG(pSer)LDNL (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.44
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 129
P value: 8.24
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.79
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: PPFIA1
Fragment: 663-672
Site: 14-3-3_pSer668
Modification: (SEP)668
Sequence: RVGSGSLDNL
Construct: RVGSG(pSer)LDNL (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.41
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 134
P value: 9.32
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.71
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: PPFIA1
Fragment: 663-672
Site: 14-3-3_pSer668
Modification: (SEP)668
Sequence: RVGSGSLDNL
Construct: biotin-ttds-RVGSG(pSer)LDNL (HPLC-purified 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): 255
Standard deviation of dissociation constant (10-6M): 35
Experiment method: competitive Fluorescence Polarization
Details of affinity fitting: Competitive binding equation in ProFit
Date of measurement: 2022-01-18
Number of measurements: 3
Measured pKd: 3.59
DOI: 10.1101/2022.12.08.519103

Experiment series 8

Protein A

Protein: PPFIA1
Fragment: 695-704
Site: 14-3-3_pSer700
Modification: (SEP)700
Sequence: PGSGRSTPRR
Construct: PGSGR(pSer)TPRR (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.36
Holdup BI standard deviation: 0.64
Immobilized partner concentration (10-6M): 134
P value: 0.27
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: PPFIA1
Fragment: 758-767
Site: 14-3-3_pSer763
Modification: (SEP)763
Sequence: ALHTVSHEDI
Construct: ALHTV(pSer)HEDI (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.48
Holdup BI standard deviation: 0.23
Immobilized partner concentration (10-6M): 134
P value: 2.04
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.83
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: PPFIA1
Fragment: 758-767
Site: 14-3-3_pSer763
Modification: (SEP)763
Sequence: ALHTVSHEDI
Construct: ALHTV(pSer)HEDI (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.11
Holdup BI standard deviation: 0.49
Immobilized partner concentration (10-6M): 129
P value: 0.17
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 11

Protein A

Protein: PPFIA1
Fragment: 801-810
Site: 14-3-3_pSer806
Modification: (SEP)806
Sequence: KGIKSSIGRL
Construct: KGIKS(pSer)IGRL (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.03
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 134
P value: 0.45
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 12

Protein A

Protein: PPFIA1
Fragment: 801-810
Site: 14-3-3_pSer806
Modification: (SEP)806
Sequence: KGIKSSIGRL
Construct: KGIKS(pSer)IGRL (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0
Holdup BI standard deviation: 0.13
Immobilized partner concentration (10-6M): 129
P value: 0
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 13

Protein A

Protein: PPFIA1
Fragment: 1154-1163
Site: 14-3-3_pThr1159
Modification: (TPO)1159
Sequence: AGSAETLPAN
Construct: AGSAE(pThr)LPAN (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.57
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 129
P value: 8.42
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.02
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: PPFIA1
Fragment: 1154-1163
Site: 14-3-3_pThr1159
Modification: (TPO)1159
Sequence: AGSAETLPAN
Construct: AGSAE(pThr)LPAN (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.55
Holdup BI standard deviation: 0.1
Immobilized partner concentration (10-6M): 134
P value: 2.63
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.96
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: PPFIA1
Fragment: 567-576
Site: 14-3-3_pThr572
Modification: (TPO)572
Sequence: PSKVQTLNEQ
Construct: PSKVQ(pThr)LNEQ (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.14
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 134
P value: 2.79
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 16

Protein A

Protein: PPFIA1
Fragment: 567-576
Site: 14-3-3_pThr572
Modification: (TPO)572
Sequence: PSKVQTLNEQ
Construct: PSKVQ(pThr)LNEQ (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.15
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 129
P value: 4.67
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 17

Protein A

Protein: PPFIA1
Fragment: 735-744
Site: 14-3-3_pThr740
Modification: (TPO)740
Sequence: TIKCETSPPS
Construct: TIKCE(pThr)SPPS (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.1
Holdup BI standard deviation: 0.87
Immobilized partner concentration (10-6M): 129
P value: 0.09
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 18

Protein A

Protein: PPFIA1
Fragment: 1134-1143
Site: 14-3-3_pSer1139
Modification: (SEP)1139
Sequence: FRRAPSWRKK
Construct: FRRAP(pSer)WRKK (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
Average holdup BI: 0.11
Holdup BI standard deviation: 0.33
Immobilized partner concentration (10-6M): 115
P value: 0.24
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: PPFIA1
Fragment: 525-534
Site: 14-3-3_pSer530
Modification: (SEP)530
Sequence: RPHLGSVPDF
Construct: RPHLG(pSer)VPDF (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
Average holdup BI: 0.64
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 115
P value: 5.83
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.21
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 20

Protein A

Protein: PPFIA1
Fragment: 663-672
Site: 14-3-3_pSer668
Modification: (SEP)668
Sequence: RVGSGSLDNL
Construct: RVGSG(pSer)LDNL (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
Average holdup BI: 0.39
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 115
P value: 5.86
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.76
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 21

Protein A

Protein: PPFIA1
Fragment: 695-704
Site: 14-3-3_pSer700
Modification: (SEP)700
Sequence: PGSGRSTPRR
Construct: PGSGR(pSer)TPRR (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
Average holdup BI: 0.71
Holdup BI standard deviation: 0.1
Immobilized partner concentration (10-6M): 115
P value: 1.53
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.34
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 22

Protein A

Protein: PPFIA1
Fragment: 758-767
Site: 14-3-3_pSer763
Modification: (SEP)763
Sequence: ALHTVSHEDI
Construct: ALHTV(pSer)HEDI (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
Average holdup BI: -0.35
Holdup BI standard deviation: 0.56
Immobilized partner concentration (10-6M): 115
P value: 0.72
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 23

Protein A

Protein: PPFIA1
Fragment: 801-810
Site: 14-3-3_pSer806
Modification: (SEP)806
Sequence: KGIKSSIGRL
Construct: KGIKS(pSer)IGRL (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
Average holdup BI: 0.12
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 115
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 24

Protein A

Protein: PPFIA1
Fragment: 1154-1163
Site: 14-3-3_pThr1159
Modification: (TPO)1159
Sequence: AGSAETLPAN
Construct: AGSAE(pThr)LPAN (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
Average holdup BI: 0.58
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 115
P value: 5.37
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.1
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 25

Protein A

Protein: PPFIA1
Fragment: 567-576
Site: 14-3-3_pThr572
Modification: (TPO)572
Sequence: PSKVQTLNEQ
Construct: PSKVQ(pThr)LNEQ (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
Average holdup BI: 0.17
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 115
P value: 2.64
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 26

Protein A

Protein: PPFIA1
Fragment: 735-744
Site: 14-3-3_pThr740
Modification: (TPO)740
Sequence: TIKCETSPPS
Construct: TIKCE(pThr)SPPS (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
Average holdup BI: -0.5
Holdup BI standard deviation: 0.56
Immobilized partner concentration (10-6M): 115
P value: 1
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))