Interaction Details


Details of experiments between P27348 and P40818
Note that the results of all experiments are listed, regardless of the modification states of the fragments.


Experiment series 1

Protein A

Protein: YWHAQ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3tau

Protein B

Protein: USP8
Fragment: 636-645
Site: 14-3-3_pSer641
Modification: (SEP)641
Sequence: LTRARSEEMG
Construct: LTRAR(pSer)EEMG (crude synthetic peptide)
Average holdup BI: 0.4
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 130
P value: 6.01
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.71
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: YWHAQ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3tau

Protein B

Protein: USP8
Fragment: 686-695
Site: 14-3-3_pSer691
Modification: (SEP)691
Sequence: SSAPPSTPPT
Construct: SSAPP(pSer)TPPT (crude synthetic peptide)
Average holdup BI: -0.04
Holdup BI standard deviation: 0.12
Immobilized partner concentration (10-6M): 130
P value: 0.32
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: YWHAQ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3tau

Protein B

Protein: USP8
Fragment: 712-724
Site: 14-3-3_pSer718
Modification: (SEP)718
Sequence: LKRSYSSPDI
Construct: KLKRSY(pSer)SPDITQ (crude synthetic peptide)
Average holdup BI: 0.95
Holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 130
P value: 9.81
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 5.17
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: YWHAQ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3tau

Protein B

Protein: USP8
Fragment: 713-722
Site: 14-3-3_pSer718
Modification: (SEP)718
Sequence: LKRSYSSPDI
Construct: LKRSY(pSer)SPDI (crude synthetic peptide)
Average holdup BI: 0.72
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 130
P value: 8.89
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))