Interaction Details


Details of experiments between P31946 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: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta

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: 0.52
Holdup BI standard deviation: 0.09
Immobilized partner concentration (10-6M): 102
P value: 4.85
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.03
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: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta

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: 0.09
Holdup BI standard deviation: 0.82
Immobilized partner concentration (10-6M): 134
P value: 0.07
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 3

Protein A

Protein: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta

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: -0.12
Holdup BI standard deviation: 0.08
Immobilized partner concentration (10-6M): 134
P value: 1.64
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 4

Protein A

Protein: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta

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: -0.07
Holdup BI standard deviation: 0.15
Immobilized partner concentration (10-6M): 102
P value: 0.44
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: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta

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: 0.96
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 102
P value: 9.84
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 5.4
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: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta

Protein B

Protein: USP8
Fragment: 712-724
Site: 14-3-3_pSer718
Modification: (SEP)718
Sequence: LKRSYSSPDI
Construct: KLKRSY(pSer)SPDITQ (crude synthetic peptide)
Measured dissociation constant (10-6M): <10
Immobilized partner concentration (10-6M): 134
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 1
Measured pKd: 5
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: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta

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: 0.85
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 134
P value: 7.95
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.61
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 8

Protein A

Protein: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta

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: 0.75
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 102
P value: 9.63
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.47
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))