Interaction Details


Details of experiments between Q86SQ0 and P61981
Note that the results of all experiments are listed, regardless of the modification states of the fragments.



Experiment series 1

Protein A

Protein: PHLDB2
Fragment: 195-204
Site: 14-3-3_pSer200
Modification: (SEP)200
Sequence: RSGAASMPSS
Construct: RSGAA(pSer)MPSS (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.91
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 129
P value: 11.4
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.88
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: PHLDB2
Fragment: 195-204
Site: 14-3-3_pSer200
Modification: (SEP)200
Sequence: RSGAASMPSS
Construct: RSGAA(pSer)MPSS (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.9
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 134
P value: 5.53
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.83
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: PHLDB2
Fragment: 207-216
Site: 14-3-3_pSer212
Modification: (SEP)212
Sequence: QARKMSIQDS
Construct: QARKM(pSer)IQDS (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.18
Holdup BI standard deviation: 0.17
Immobilized partner concentration (10-6M): 134
P value: 0.9
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: PHLDB2
Fragment: 207-216
Site: 14-3-3_pSer212
Modification: (SEP)212
Sequence: QARKMSIQDS
Construct: QARKM(pSer)IQDS (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.1
Holdup BI standard deviation: 0.2
Immobilized partner concentration (10-6M): 129
P value: 0.46
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: PHLDB2
Fragment: 237-246
Site: 14-3-3_pSer242
Modification: (SEP)242
Sequence: RTRKYSSSSL
Construct: RTRKY(pSer)SSSL (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.28
Holdup BI standard deviation: 0.24
Immobilized partner concentration (10-6M): 134
P value: 0.81
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 6

Protein A

Protein: PHLDB2
Fragment: 237-246
Site: 14-3-3_pSer242
Modification: (SEP)242
Sequence: RTRKYSSSSL
Construct: RTRKY(pSer)SSSL (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.25
Holdup BI standard deviation: 0.27
Immobilized partner concentration (10-6M): 129
P value: 0.87
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 7

Protein A

Protein: PHLDB2
Fragment: 272-281
Site: 14-3-3_pSer277
Modification: (SEP)277
Sequence: LPPRNSLGNS
Construct: LPPRN(pSer)LGNS (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.19
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 134
P value: 5.2
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 8

Protein A

Protein: PHLDB2
Fragment: 272-281
Site: 14-3-3_pSer277
Modification: (SEP)277
Sequence: LPPRNSLGNS
Construct: LPPRN(pSer)LGNS (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.17
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 129
P value: 5.04
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 9

Protein A

Protein: PHLDB2
Fragment: 379-388
Site: 14-3-3_pSer384
Modification: (SEP)384
Sequence: TRRNFSCGSV
Construct: TRRNF(pSer)CGSV (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.21
Holdup BI standard deviation: 0.67
Immobilized partner concentration (10-6M): 134
P value: 0.21
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: PHLDB2
Fragment: 410-419
Site: 14-3-3_pSer415
Modification: (SEP)415
Sequence: RERKSSISSI
Construct: RERKS(pSer)ISSI (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.14
Holdup BI standard deviation: 0.66
Immobilized partner concentration (10-6M): 134
P value: 0.16
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 11

Protein A

Protein: PHLDB2
Fragment: 410-419
Site: 14-3-3_pSer415
Modification: (SEP)415
Sequence: RERKSSISSI
Construct: RERKS(pSer)ISSI (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.14
Holdup BI standard deviation: 0.15
Immobilized partner concentration (10-6M): 129
P value: 0.77
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 12

Protein A

Protein: PHLDB2
Fragment: 463-472
Site: 14-3-3_pSer468
Modification: (SEP)468
Sequence: PDSRLSTGTT
Construct: PDSRL(pSer)TGTT (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: -0.03
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 134
P value: 0.87
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 13

Protein A

Protein: PHLDB2
Fragment: 463-472
Site: 14-3-3_pSer468
Modification: (SEP)468
Sequence: PDSRLSTGTT
Construct: PDSRL(pSer)TGTT (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.02
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 129
P value: 0.46
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 14

Protein A

Protein: PHLDB2
Fragment: 484-493
Site: 14-3-3_pSer489
Modification: (SEP)489
Sequence: EKLQLSDEES
Construct: EKLQL(pSer)DEES (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: -0.05
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 134
P value: 1.37
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: PHLDB2
Fragment: 484-493
Site: 14-3-3_pSer489
Modification: (SEP)489
Sequence: EKLQLSDEES
Construct: EKLQL(pSer)DEES (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: -0.11
Holdup BI standard deviation: 0.12
Immobilized partner concentration (10-6M): 129
P value: 1.18
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 16

Protein A

Protein: PHLDB2
Fragment: 1123-1132
Site: 14-3-3_pThr1128
Modification: (TPO)1128
Sequence: RSHVETAGHN
Construct: RSHVE(pThr)AGHN (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: -0.12
Holdup BI standard deviation: 0.37
Immobilized partner concentration (10-6M): 129
P value: 0.2
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 17

Protein A

Protein: PHLDB2
Fragment: 893-902
Site: 14-3-3_pThr898
Modification: (TPO)898
Sequence: LYLSDTLPRK
Construct: LYLSD(pThr)LPRK (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.06
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 134
P value: 0.95
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 18

Protein A

Protein: PHLDB2
Fragment: 893-902
Site: 14-3-3_pThr898
Modification: (TPO)898
Sequence: LYLSDTLPRK
Construct: LYLSD(pThr)LPRK (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.27
Holdup BI standard deviation: 0.1
Immobilized partner concentration (10-6M): 129
P value: 2.8
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 19

Protein A

Protein: PHLDB2
Fragment: 195-204
Site: 14-3-3_pSer200
Modification: (SEP)200
Sequence: RSGAASMPSS
Construct: RSGAA(pSer)MPSS (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.85
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 115
P value: 4.22
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.7
DOI: 10.1101/2022.12.08.519103
Experimental Details

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


Experiment series 20

Protein A

Protein: PHLDB2
Fragment: 207-216
Site: 14-3-3_pSer212
Modification: (SEP)212
Sequence: QARKMSIQDS
Construct: QARKM(pSer)IQDS (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.13
Holdup BI standard deviation: 0.33
Immobilized partner concentration (10-6M): 115
P value: 0.31
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 21

Protein A

Protein: PHLDB2
Fragment: 272-281
Site: 14-3-3_pSer277
Modification: (SEP)277
Sequence: LPPRNSLGNS
Construct: LPPRN(pSer)LGNS (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.16
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 115
P value: 5.83
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 22

Protein A

Protein: PHLDB2
Fragment: 379-388
Site: 14-3-3_pSer384
Modification: (SEP)384
Sequence: TRRNFSCGSV
Construct: TRRNF(pSer)CGSV (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.2
Holdup BI standard deviation: 0.09
Immobilized partner concentration (10-6M): 115
P value: 1.93
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 23

Protein A

Protein: PHLDB2
Fragment: 463-472
Site: 14-3-3_pSer468
Modification: (SEP)468
Sequence: PDSRLSTGTT
Construct: PDSRL(pSer)TGTT (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 115
P value: 0.02
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 24

Protein A

Protein: PHLDB2
Fragment: 484-493
Site: 14-3-3_pSer489
Modification: (SEP)489
Sequence: EKLQLSDEES
Construct: EKLQL(pSer)DEES (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: -0.03
Holdup BI standard deviation: 0.09
Immobilized partner concentration (10-6M): 115
P value: 0.27
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 25

Protein A

Protein: PHLDB2
Fragment: 893-902
Site: 14-3-3_pThr898
Modification: (TPO)898
Sequence: LYLSDTLPRK
Construct: LYLSD(pThr)LPRK (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.42
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 115
P value: 4.3
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.81
DOI: 10.1101/2022.12.08.519103
Experimental Details

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