Interaction Details


Details of experiments between Q8TF72 and P31946
Note that the results of all experiments are listed, regardless of the modification states of the fragments.



Experiment series 1

Protein A

Protein: SHROOM3
Fragment: 1016-1025
Site: 14-3-3_pSer1021
Modification: (SEP)1021
Sequence: KKRSYSEPEK
Construct: KKRSY(pSer)EPEK (crude synthetic peptide)

Protein B

Protein: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.04
Holdup BI standard deviation: 0.23
Immobilized partner concentration (10-6M): 134
P value: 0.13
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 2

Protein A

Protein: SHROOM3
Fragment: 1016-1025
Site: 14-3-3_pSer1021
Modification: (SEP)1021
Sequence: KKRSYSEPEK
Construct: KKRSY(pSer)EPEK (crude synthetic peptide)

Protein B

Protein: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.04
Holdup BI standard deviation: 0.13
Immobilized partner concentration (10-6M): 102
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 3

Protein A

Protein: SHROOM3
Fragment: 1216-1225
Site: 14-3-3_pSer1221
Modification: (SEP)1221
Sequence: AGKSMSAEDL
Construct: AGKSM(pSer)AEDL (crude synthetic peptide)

Protein B

Protein: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.44
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 134
P value: 3.88
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.77
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: SHROOM3
Fragment: 1216-1225
Site: 14-3-3_pSer1221
Modification: (SEP)1221
Sequence: AGKSMSAEDL
Construct: AGKSM(pSer)AEDL (crude synthetic peptide)

Protein B

Protein: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.34
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 102
P value: 4.97
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.7
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: SHROOM3
Fragment: 1416-1425
Site: 14-3-3_pSer1421
Modification: (SEP)1421
Sequence: TRKRVSLPQW
Construct: TRKRV(pSer)LPQW (crude synthetic peptide)

Protein B

Protein: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.85
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 102
P value: 4.4
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.73
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: SHROOM3
Fragment: 1436-1445
Site: 14-3-3_pSer1441
Modification: (SEP)1441
Sequence: AAREDSLPEE
Construct: AARED(pSer)LPEE (crude synthetic peptide)

Protein B

Protein: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: -0.04
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 134
P value: 1.43
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 7

Protein A

Protein: SHROOM3
Fragment: 1436-1445
Site: 14-3-3_pSer1441
Modification: (SEP)1441
Sequence: AAREDSLPEE
Construct: AARED(pSer)LPEE (crude synthetic peptide)

Protein B

Protein: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: -0.12
Holdup BI standard deviation: 0.14
Immobilized partner concentration (10-6M): 102
P value: 1.2
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 8

Protein A

Protein: SHROOM3
Fragment: 1457-1466
Site: 14-3-3_pSer1462
Modification: (SEP)1462
Sequence: YQKQQSLPSL
Construct: YQKQQ(pSer)LPSL (crude synthetic peptide)

Protein B

Protein: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.79
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 102
P value: 6.6
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.58
DOI: 10.1101/2022.12.08.519103
Experimental Details

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


Experiment series 9

Protein A

Protein: SHROOM3
Fragment: 1457-1466
Site: 14-3-3_pSer1462
Modification: (SEP)1462
Sequence: YQKQQSLPSL
Construct: YQKQQ(pSer)LPSL (crude synthetic peptide)

Protein B

Protein: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.06
Holdup BI standard deviation: 0.09
Immobilized partner concentration (10-6M): 134
P value: 0.62
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 10

Protein A

Protein: SHROOM3
Fragment: 414-423
Site: 14-3-3_pSer419
Modification: (SEP)419
Sequence: RPQANSLGSL
Construct: RPQAN(pSer)LGSL (crude synthetic peptide)

Protein B

Protein: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.68
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 102
P value: 7.86
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.32
DOI: 10.1101/2022.12.08.519103
Experimental Details

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


Experiment series 11

Protein A

Protein: SHROOM3
Fragment: 414-423
Site: 14-3-3_pSer419
Modification: (SEP)419
Sequence: RPQANSLGSL
Construct: RPQAN(pSer)LGSL (crude synthetic peptide)

Protein B

Protein: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.68
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 134
P value: 7.76
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.2
DOI: 10.1101/2022.12.08.519103
Experimental Details

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


Experiment series 12

Protein A

Protein: SHROOM3
Fragment: 618-627
Site: 14-3-3_pSer623
Modification: (SEP)623
Sequence: RSSRLSEPWE
Construct: RSSRL(pSer)EPWE (crude synthetic peptide)

Protein B

Protein: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.54
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 102
P value: 7.38
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.06
DOI: 10.1101/2022.12.08.519103
Experimental Details

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


Experiment series 13

Protein A

Protein: SHROOM3
Fragment: 618-627
Site: 14-3-3_pSer623
Modification: (SEP)623
Sequence: RSSRLSEPWE
Construct: RSSRL(pSer)EPWE (crude synthetic peptide)

Protein B

Protein: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.56
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 134
P value: 7.73
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.98
DOI: 10.1101/2022.12.08.519103
Experimental Details

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


Experiment series 14

Protein A

Protein: SHROOM3
Fragment: 716-725
Site: 14-3-3_pSer721
Modification: (SEP)721
Sequence: LDRQVSYPRP
Construct: LDRQV(pSer)YPRP (crude synthetic peptide)

Protein B

Protein: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: -0.04
Holdup BI standard deviation: 0.33
Immobilized partner concentration (10-6M): 134
P value: 0.09
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 15

Protein A

Protein: SHROOM3
Fragment: 716-725
Site: 14-3-3_pSer721
Modification: (SEP)721
Sequence: LDRQVSYPRP
Construct: LDRQV(pSer)YPRP (crude synthetic peptide)

Protein B

Protein: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.18
Holdup BI standard deviation: 0.17
Immobilized partner concentration (10-6M): 102
P value: 1.17
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 16

Protein A

Protein: SHROOM3
Fragment: 949-958
Site: 14-3-3_pSer954
Modification: (SEP)954
Sequence: GAPVASRSWR
Construct: GAPVA(pSer)RSWR (crude synthetic peptide)

Protein B

Protein: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.06
Holdup BI standard deviation: 0.08
Immobilized partner concentration (10-6M): 134
P value: 0.65
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 17

Protein A

Protein: SHROOM3
Fragment: 949-958
Site: 14-3-3_pSer954
Modification: (SEP)954
Sequence: GAPVASRSWR
Construct: GAPVA(pSer)RSWR (crude synthetic peptide)

Protein B

Protein: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: -0.03
Holdup BI standard deviation: 0.09
Immobilized partner concentration (10-6M): 102
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 18

Protein A

Protein: SHROOM3
Fragment: 107-116
Site: 14-3-3_pThr112
Modification: (TPO)112
Sequence: RRDVCTDPGH
Construct: RRDVC(pThr)DPGH (crude synthetic peptide)

Protein B

Protein: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: -0.4
Holdup BI standard deviation: 0.62
Immobilized partner concentration (10-6M): 134
P value: 0.89
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 19

Protein A

Protein: SHROOM3
Fragment: 107-116
Site: 14-3-3_pThr112
Modification: (TPO)112
Sequence: RRDVCTDPGH
Construct: RRDVC(pThr)DPGH (crude synthetic peptide)

Protein B

Protein: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: -0.06
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 102
P value: 0.94
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))