Interaction Details

Details of experiments between P15056 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: BRAF
Fragment: 334-343
Site: 14-3-3_pSer339
Modification: (SEP)339
Sequence: PSPSKSIPIP
Construct: PSPSK(pSer)IPIP (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: 0.39

Holdup BI standard deviation: 0.06

Immobilized partner concentration (10^-6M): 102

P value: 5

Experiment method: HU Multiplex

Details of affinity fitting: hyperbolic binding equation

Date of measurement: 2021-10-13

Number of measurements: 5

Measured pK_d: 3.8

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: BRAF
Fragment: 334-343
Site: 14-3-3_pSer339
Modification: (SEP)339
Sequence: PSPSKSIPIP
Construct: PSPSK(pSer)IPIP (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: 0.04

Immobilized partner concentration (10^-6M): 134

P value: 4.56

Experiment method: HU Multiplex

Details of affinity fitting: hyperbolic binding equation

Date of measurement: 2022-02-17

Number of measurements: 4

Measured pK_d: 3.7

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: BRAF
Fragment: 360-369
Site: 14-3-3_pSer365
Modification: (SEP)365
Sequence: RDRSSSAPNV
Construct: RDRSS(pSer)APNV (crude synthetic peptide)

Protein B

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

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 pK_d: 5

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: BRAF
Fragment: 389-398
Site: 14-3-3_pSer394
Modification: (SEP)394
Sequence: RGDGGSTTGL
Construct: RGDGG(pSer)TTGL (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: -0.09

Holdup BI standard deviation: 0.04

Immobilized partner concentration (10^-6M): 134

P value: 2.62

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 5

Protein A

Protein: BRAF
Fragment: 389-398
Site: 14-3-3_pSer394
Modification: (SEP)394
Sequence: RGDGGSTTGL
Construct: RGDGG(pSer)TTGL (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: -0.07

Holdup BI standard deviation: 0.08

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))

Experiment series 6

Protein A

Protein: BRAF
Fragment: 441-450
Site: 14-3-3_pSer446
Modification: (SEP)446
Sequence: LGRRDSSDDW
Construct: LGRRD(pSer)SDDW (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: 0.05

Immobilized partner concentration (10^-6M): 134

P value: 1.09

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 7

Protein A

Protein: BRAF
Fragment: 441-450
Site: 14-3-3_pSer446
Modification: (SEP)446
Sequence: LGRRDSSDDW
Construct: LGRRD(pSer)SDDW (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: 0.07

Immobilized partner concentration (10^-6M): 102

P value: 0.62

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: BRAF
Fragment: 462-471
Site: 14-3-3_pSer467
Modification: (SEP)467
Sequence: RIGSGSFGTV
Construct: RIGSG(pSer)FGTV (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: 0.17

Holdup BI standard deviation: 0.03

Immobilized partner concentration (10^-6M): 134

P value: 4.51

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 9

Protein A

Protein: BRAF
Fragment: 462-471
Site: 14-3-3_pSer467
Modification: (SEP)467
Sequence: RIGSGSFGTV
Construct: RIGSG(pSer)FGTV (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: 0.17

Holdup BI standard deviation: 0.1

Immobilized partner concentration (10^-6M): 102

P value: 1.87

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 10

Protein A

Protein: BRAF
Fragment: 574-583
Site: 14-3-3_pSer579
Modification: (SEP)579
Sequence: HRDLKSNNIF
Construct: HRDLK(pSer)NNIF (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: 0.22

Holdup BI standard deviation: 0.39

Immobilized partner concentration (10^-6M): 134

P value: 0.4

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: BRAF
Fragment: 574-583
Site: 14-3-3_pSer579
Modification: (SEP)579
Sequence: HRDLKSNNIF
Construct: HRDLK(pSer)NNIF (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: 0.11

Holdup BI standard deviation: 0.41

Immobilized partner concentration (10^-6M): 102

P value: 0.22

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: BRAF
Fragment: 597-606
Site: 14-3-3_pSer602
Modification: (SEP)602
Sequence: LATVKSRWSG
Construct: LATVK(pSer)RWSG (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: 0.1

Holdup BI standard deviation: 0.06

Immobilized partner concentration (10^-6M): 134

P value: 1.71

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 13

Protein A

Protein: BRAF
Fragment: 597-606
Site: 14-3-3_pSer602
Modification: (SEP)602
Sequence: LATVKSRWSG
Construct: LATVK(pSer)RWSG (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: -0.02

Holdup BI standard deviation: 0.06

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 14

Protein A

Protein: BRAF
Fragment: 609-618
Site: 14-3-3_pSer614
Modification: (SEP)614
Sequence: QFEQLSGSIL
Construct: QFEQL(pSer)GSIL (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: 0.13

Immobilized partner concentration (10^-6M): 134

P value: 0.23

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: BRAF
Fragment: 609-618
Site: 14-3-3_pSer614
Modification: (SEP)614
Sequence: QFEQLSGSIL
Construct: QFEQL(pSer)GSIL (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

Holdup BI standard deviation: 0.39

Immobilized partner concentration (10^-6M): 102

P value: 0

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: BRAF
Fragment: 724-733
Site: 14-3-3_pSer729
Modification: (SEP)729
Sequence: IHRSASEPSL
Construct: IHRSA(pSer)EPSL (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: 0.92

Holdup BI standard deviation: 0.03

Immobilized partner concentration (10^-6M): 102

P value: 8.23

Experiment method: HU Multiplex

Details of affinity fitting: hyperbolic binding equation

Date of measurement: 2021-10-13

Number of measurements: 5

Measured pK_d: 5.04

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: BRAF
Fragment: 724-733
Site: 14-3-3_pSer729
Modification: (SEP)729
Sequence: IHRSASEPSL
Construct: IHRSA(pSer)EPSL (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: 0.82

Holdup BI standard deviation: 0.24

Immobilized partner concentration (10^-6M): 134

P value: 1.55

Experiment method: HU Multiplex

Details of affinity fitting: hyperbolic binding equation

Date of measurement: 2022-02-17

Number of measurements: 4

Measured pK_d: 4.52

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: BRAF
Fragment: 465-474
Site: 14-3-3_pThr470
Modification: (TPO)470
Sequence: SGSFGTVYKG
Construct: SGSFG(pThr)VYKG (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: 0.37

Holdup BI standard deviation: 0.06

Immobilized partner concentration (10^-6M): 102

P value: 5.12

Experiment method: HU Multiplex

Details of affinity fitting: hyperbolic binding equation

Date of measurement: 2021-10-13

Number of measurements: 5

Measured pK_d: 3.77

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: BRAF
Fragment: 465-474
Site: 14-3-3_pThr470
Modification: (TPO)470
Sequence: SGSFGTVYKG
Construct: SGSFG(pThr)VYKG (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: 0.36

Holdup BI standard deviation: 0.02

Immobilized partner concentration (10^-6M): 134

P value: 6.45

Experiment method: HU Multiplex

Details of affinity fitting: hyperbolic binding equation

Date of measurement: 2022-02-17

Number of measurements: 4

Measured pK_d: 3.63

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: BRAF
Fragment: 594-603
Site: 14-3-3_pThr599
Modification: (TPO)599
Sequence: DFGLATVKSR
Construct: DFGLA(pThr)VKSR (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: 0.09

Holdup BI standard deviation: 0.06

Immobilized partner concentration (10^-6M): 134

P value: 1.49

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: BRAF
Fragment: 594-603
Site: 14-3-3_pThr599
Modification: (TPO)599
Sequence: DFGLATVKSR
Construct: DFGLA(pThr)VKSR (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: 0.01

Holdup BI standard deviation: 0.06

Immobilized partner concentration (10^-6M): 102

P value: 0.18

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))