Interaction Details


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

Protein B

Protein: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.14
Holdup BI standard deviation: 0.62
Immobilized partner concentration (10-6M): 134
P value: 0.17
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: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: -0.03
Holdup BI standard deviation: 0.55
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 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: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.71
Holdup BI standard deviation: 0.23
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
Measured pKd: 4.26
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: PPFIA1
Fragment: 525-534
Site: 14-3-3_pSer530
Modification: (SEP)530
Sequence: RPHLGSVPDF
Construct: RPHLG(pSer)VPDF (crude synthetic peptide)

Protein B

Protein: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.57
Holdup BI standard deviation: 0.1
Immobilized partner concentration (10-6M): 102
P value: 3.68
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.12
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: PPFIA1
Fragment: 663-672
Site: 14-3-3_pSer668
Modification: (SEP)668
Sequence: RVGSGSLDNL
Construct: RVGSG(pSer)LDNL (crude synthetic peptide)

Protein B

Protein: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.35
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 102
P value: 5.24
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.72
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: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.33
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 134
P value: 8.45
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.57
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: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Measured dissociation constant (10-6M): 497
Standard deviation of dissociation constant (10-6M): 125
Experiment method: competitive Fluorescence Polarization
Details of affinity fitting: Competitive binding equation in ProFit
Date of measurement: 2022-02-08
Number of measurements: 3
Measured pKd: 3.3
DOI: 10.1101/2022.12.08.519103

Experiment series 8

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: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.32
Holdup BI standard deviation: 0.1
Immobilized partner concentration (10-6M): 134
P value: 2.56
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.55
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: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: -0.11
Holdup BI standard deviation: 0.53
Immobilized partner concentration (10-6M): 102
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))

Experiment series 10

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: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.17
Holdup BI standard deviation: 0.21
Immobilized partner concentration (10-6M): 134
P value: 0.81
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 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: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0
Holdup BI standard deviation: 0.12
Immobilized partner concentration (10-6M): 102
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 12

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: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.45
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 102
P value: 6
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.9
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: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.38
Holdup BI standard deviation: 0.13
Immobilized partner concentration (10-6M): 134
P value: 1.92
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.67
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 14

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: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.07
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 134
P value: 1.05
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: PPFIA1
Fragment: 567-576
Site: 14-3-3_pThr572
Modification: (TPO)572
Sequence: PSKVQTLNEQ
Construct: PSKVQ(pThr)LNEQ (crude synthetic peptide)

Protein B

Protein: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.08
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 102
P value: 2.66
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))