Interaction Details


Details of experiments between Q13136 and Q04917
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: 525-534
Site: 14-3-3_pSer530
Modification: (SEP)530
Sequence: RPHLGSVPDF
Construct: RPHLG(pSer)VPDF (crude synthetic peptide)

Protein B

Protein: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta
Average holdup BI: 0.56
Holdup BI standard deviation: 0.09
Immobilized partner concentration (10-6M): 130
P value: 3.64
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.98
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: PPFIA1
Fragment: 663-672
Site: 14-3-3_pSer668
Modification: (SEP)668
Sequence: RVGSGSLDNL
Construct: RVGSG(pSer)LDNL (crude synthetic peptide)

Protein B

Protein: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta
Average holdup BI: 0.36
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 130
P value: 7.16
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.63
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: 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: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta
Measured dissociation constant (10-6M): 324
Standard deviation of dissociation constant (10-6M): 24
Experiment method: competitive Fluorescence Polarization
Details of affinity fitting: Competitive binding equation in ProFit
Date of measurement: 2022-01-19
Number of measurements: 3
Measured pKd: 3.49
DOI: 10.1101/2022.12.08.519103

Experiment series 4

Protein A

Protein: PPFIA1
Fragment: 692-701
Site: 14-3-3_pSer697
Modification: (SEP)697
Sequence: SSPPGSGRST
Construct: SSPPG(pSer)GRST (crude synthetic peptide)

Protein B

Protein: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta
Average holdup BI: -0.44
Holdup BI standard deviation: 1.07
Immobilized partner concentration (10-6M): 130
P value: 0.63
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: PPFIA1
Fragment: 758-767
Site: 14-3-3_pSer763
Modification: (SEP)763
Sequence: ALHTVSHEDI
Construct: ALHTV(pSer)HEDI (crude synthetic peptide)

Protein B

Protein: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta
Average holdup BI: -0.15
Holdup BI standard deviation: 0.53
Immobilized partner concentration (10-6M): 130
P value: 0.3
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 6

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: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta
Average holdup BI: 0.11
Holdup BI standard deviation: 0.2
Immobilized partner concentration (10-6M): 130
P value: 0.52
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 7

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: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta
Average holdup BI: 0.49
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 130
P value: 7.06
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.87
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: PPFIA1
Fragment: 567-576
Site: 14-3-3_pThr572
Modification: (TPO)572
Sequence: PSKVQTLNEQ
Construct: PSKVQ(pThr)LNEQ (crude synthetic peptide)

Protein B

Protein: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta
Average holdup BI: 0.11
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 130
P value: 3.55
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))