Interaction Details

Details of experiments between Q86SQ0 and P62258
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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Average holdup BI: 0.75

Holdup BI standard deviation: 0.06

Immobilized partner concentration (10^-6M): 116

P value: 4.79

Experiment method: HU Multiplex

Details of affinity fitting: hyperbolic binding equation

Date of measurement: 2022-02-17

Number of measurements: 4

Measured pK_d: 4.41

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

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Average holdup BI: 0.74

Holdup BI standard deviation: 0.02

Immobilized partner concentration (10^-6M): 143

P value: 9.73

Experiment method: HU Multiplex

Details of affinity fitting: hyperbolic binding equation

Date of measurement: 2021-10-13

Number of measurements: 5

Measured pK_d: 4.3

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

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Average holdup BI: 0.15

Holdup BI standard deviation: 0.22

Immobilized partner concentration (10^-6M): 116

P value: 0.57

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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Average holdup BI: -0.03

Holdup BI standard deviation: 0.18

Immobilized partner concentration (10^-6M): 143

P value: 0.15

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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Average holdup BI: -0.01

Holdup BI standard deviation: 0.75

Immobilized partner concentration (10^-6M): 116

P value: 0.01

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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Average holdup BI: -0.28

Holdup BI standard deviation: 0.66

Immobilized partner concentration (10^-6M): 143

P value: 0.41

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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Average holdup BI: 0.06

Holdup BI standard deviation: 0.03

Immobilized partner concentration (10^-6M): 116

P value: 2.04

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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Average holdup BI: 0.04

Holdup BI standard deviation: 0.05

Immobilized partner concentration (10^-6M): 143

P value: 1.06

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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Average holdup BI: 0

Holdup BI standard deviation: 0.82

Immobilized partner concentration (10^-6M): 116

P value: 0

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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Average holdup BI: -0.08

Holdup BI standard deviation: 0.81

Immobilized partner concentration (10^-6M): 116

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 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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Average holdup BI: 0.1

Holdup BI standard deviation: 0.14

Immobilized partner concentration (10^-6M): 143

P value: 0.57

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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Average holdup BI: -0.06

Holdup BI standard deviation: 0.05

Immobilized partner concentration (10^-6M): 116

P value: 1.17

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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Average holdup BI: 0.03

Holdup BI standard deviation: 0.04

Immobilized partner concentration (10^-6M): 143

P value: 0.65

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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Average holdup BI: 0.05

Holdup BI standard deviation: 0.13

Immobilized partner concentration (10^-6M): 116

P value: 0.3

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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Average holdup BI: -0.02

Holdup BI standard deviation: 0.14

Immobilized partner concentration (10^-6M): 143

P value: 0.1

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

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Average holdup BI: -0.09

Holdup BI standard deviation: 0.13

Immobilized partner concentration (10^-6M): 116

P value: 0.71

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

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Average holdup BI: 0.04

Holdup BI standard deviation: 0.09

Immobilized partner concentration (10^-6M): 143

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))