Interaction Details


Details of experiments between Q4KWH8 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: PLCH1
Fragment: 1411-1420
Site: 14-3-3_pSer1416
Modification: (SEP)1416
Sequence: ETLRPSVPEI
Construct: ETLRP(pSer)VPEI (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: 0.05
Holdup BI standard deviation: 0.22
Immobilized partner concentration (10-6M): 130
P value: 0.18
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: PLCH1
Fragment: 918-927
Site: 14-3-3_pSer923
Modification: (SEP)923
Sequence: LRRTASAPAK
Construct: LRRTA(pSer)APAK (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: 0.14
Holdup BI standard deviation: 0.14
Immobilized partner concentration (10-6M): 130
P value: 1.07
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: PLCH1
Fragment: 965-974
Site: 14-3-3_pSer970
Modification: (SEP)970
Sequence: QARPVSMPVD
Construct: QARPV(pSer)MPVD (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: 0.84
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 130
P value: 7.35
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.6
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: PLCH1
Fragment: 1315-1324
Site: 14-3-3_pThr1320
Modification: (TPO)1320
Sequence: LPKSPTKGED
Construct: LPKSP(pThr)KGED (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: 0.01
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 130
P value: 0.27
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: PLCH1
Fragment: 1515-1524
Site: 14-3-3_pThr1520
Modification: (TPO)1520
Sequence: SKSFVTTGIR
Construct: SKSFV(pThr)TGIR (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: 0.06
Immobilized partner concentration (10-6M): 130
P value: 4.86
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.86
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: PLCH1
Fragment: 1515-1524
Site: 14-3-3_pThr1520
Modification: (TPO)1520
Sequence: SKSFVTTGIR
Construct: biotin-ttds-SKSFV(pThr)TGIR (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): 162
Standard deviation of dissociation constant (10-6M): 12
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: 3.79
DOI: 10.1101/2022.12.08.519103