Interaction Details


Details of experiments between P15056 and P31947
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.27
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 142
P value: 4.72
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.42
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.22
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 113
P value: 2.72
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.38
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.88
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 142
P value: 2.01
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.7
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 4

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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.78
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 113
P value: 3.6
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.48
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: -0.03
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 113
P value: 0.58
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: BRAF
Fragment: 389-398
Site: 14-3-3_pSer394
Modification: (SEP)394
Sequence: RGDGGSTTGL
Construct: RGDGG(pSer)TTGL (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: -0.02
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 142
P value: 0.24
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: BRAF
Fragment: 441-450
Site: 14-3-3_pSer446
Modification: (SEP)446
Sequence: LGRRDSSDDW
Construct: LGRRD(pSer)SDDW (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.06
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 113
P value: 1.03
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: BRAF
Fragment: 441-450
Site: 14-3-3_pSer446
Modification: (SEP)446
Sequence: LGRRDSSDDW
Construct: LGRRD(pSer)SDDW (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: -0.03
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 142
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 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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.07
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 113
P value: 2.06
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: BRAF
Fragment: 462-471
Site: 14-3-3_pSer467
Modification: (SEP)467
Sequence: RIGSGSFGTV
Construct: RIGSG(pSer)FGTV (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.08
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 142
P value: 1.73
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 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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: -0.12
Holdup BI standard deviation: 0.54
Immobilized partner concentration (10-6M): 113
P value: 0.2
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 12

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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: -0.06
Holdup BI standard deviation: 0.48
Immobilized partner concentration (10-6M): 142
P value: 0.11
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 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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.04
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 113
P value: 1.26
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 14

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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: -0.03
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 142
P value: 0.33
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 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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.06
Holdup BI standard deviation: 0.14
Immobilized partner concentration (10-6M): 113
P value: 0.33
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 16

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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: -0.4
Holdup BI standard deviation: 0.42
Immobilized partner concentration (10-6M): 142
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 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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.73
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 142
P value: 7.39
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.28
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: BRAF
Fragment: 465-474
Site: 14-3-3_pThr470
Modification: (TPO)470
Sequence: SGSFGTVYKG
Construct: SGSFG(pThr)VYKG (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.14
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 113
P value: 3.74
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: BRAF
Fragment: 465-474
Site: 14-3-3_pThr470
Modification: (TPO)470
Sequence: SGSFGTVYKG
Construct: SGSFG(pThr)VYKG (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.19
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 142
P value: 4.45
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 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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.02
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 113
P value: 0.18
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.04
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 142
P value: 0.67
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 22

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: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.37
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 111
P value: 6.7
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.73
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: BRAF
Fragment: 360-369
Site: 14-3-3_pSer365
Modification: (SEP)365
Sequence: RDRSSSAPNV
Construct: RDRSS(pSer)APNV (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: -0.1
Holdup BI standard deviation: 0.74
Immobilized partner concentration (10-6M): 111
P value: 0.11
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: BRAF
Fragment: 389-398
Site: 14-3-3_pSer394
Modification: (SEP)394
Sequence: RGDGGSTTGL
Construct: RGDGG(pSer)TTGL (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.08
Holdup BI standard deviation: 0.51
Immobilized partner concentration (10-6M): 111
P value: 0.11
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: BRAF
Fragment: 441-450
Site: 14-3-3_pSer446
Modification: (SEP)446
Sequence: LGRRDSSDDW
Construct: LGRRD(pSer)SDDW (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: -0.01
Holdup BI standard deviation: 0.08
Immobilized partner concentration (10-6M): 111
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 26

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: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.22
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 111
P value: 3.57
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.41
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 27

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: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.19
Holdup BI standard deviation: 0.09
Immobilized partner concentration (10-6M): 111
P value: 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 28

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: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.02
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 111
P value: 0.26
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 29

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: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: -0.07
Holdup BI standard deviation: 0.08
Immobilized partner concentration (10-6M): 111
P value: 0.86
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 30

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: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.61
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 111
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
Measured pKd: 4.15
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 31

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: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.13
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 111
P value: 3.04
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 32

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: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.01
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 111
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))