Interaction Details


Details of experiments between Q9Y2J2 and P63104
Note that the results of all experiments are listed, regardless of the modification states of the fragments.


Experiment series 1

Protein A

Protein: EPB41L3
Fragment: 415-424
Site: 14-3-3_pSer420
Modification: (SEP)420
Sequence: QTRRASALID
Construct: QTRRA(pSer)ALID (crude synthetic peptide)

Protein B

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta
Average holdup BI: 0.1
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 132
P value: 2.71
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 2

Protein A

Protein: EPB41L3
Fragment: 438-447
Site: 14-3-3_pSer443
Modification: (SEP)443
Sequence: YTMSRSLDGA
Construct: YTMSR(pSer)LDGA (crude synthetic peptide)

Protein B

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta
Average holdup BI: 0.76
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 132
P value: 8.6
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.38
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: EPB41L3
Fragment: 455-464
Site: 14-3-3_pSer460
Modification: (SEP)460
Sequence: TTKGISQTNL
Construct: TTKGI(pSer)QTNL (crude synthetic peptide)

Protein B

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta
Average holdup BI: 0.04
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 132
P value: 1.97
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 4

Protein A

Protein: EPB41L3
Fragment: 952-961
Site: 14-3-3_pSer957
Modification: (SEP)957
Sequence: KTETISFGSV
Construct: KTETI(pSer)FGSV (crude synthetic peptide)

Protein B

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta
Average holdup BI: 0.11
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 132
P value: 2.19
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: EPB41L3
Fragment: 396-405
Site: 14-3-3_pThr401
Modification: (TPO)401
Sequence: PKKFLTLGSK
Construct: biotin-ttds-PKKFL(pThr)LGSK (HPLC-purified synthetic peptide)

Protein B

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta
Measured dissociation constant (10-6M): 20
Standard deviation of dissociation constant (10-6M): 0.5
Experiment method: competitive Fluorescence Polarization
Details of affinity fitting: Competitive binding equation in ProFit
Date of measurement: 2022-02-10
Number of measurements: 3
Measured pKd: 4.71
DOI: 10.1101/2022.12.08.519103

Experiment series 6

Protein A

Protein: EPB41L3
Fragment: 396-405
Site: 14-3-3_pThr401
Modification: (TPO)401
Sequence: PKKFLTLGSK
Construct: biotin-ttds-PKKFL(pThr)LGSK (HPLC-purified synthetic peptide)

Protein B

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta
Measured dissociation constant (10-6M): 30
Standard deviation of dissociation constant (10-6M): 0.6
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: 4.52
DOI: 10.1101/2022.12.08.519103

Experiment series 7

Protein A

Protein: EPB41L3
Fragment: 396-405
Site: 14-3-3_pThr401
Modification: (TPO)401
Sequence: PKKFLTLGSK
Construct: PKKFL(pThr)LGSK (crude synthetic peptide)

Protein B

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta
Average holdup BI: 0.73
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 132
P value: 7.82
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.31
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: EPB41L3
Fragment: 756-765
Site: 14-3-3_pThr761
Modification: (TPO)761
Sequence: EKRLSTSPVR
Construct: EKRLS(pThr)SPVR (crude synthetic peptide)

Protein B

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta
Average holdup BI: -0.27
Holdup BI standard deviation: 1.13
Immobilized partner concentration (10-6M): 132
P value: 0.21
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))