Interaction Details


Details of experiments between P63104 and P15056
Note that the results of all experiments are listed, regardless of the modification states of the fragments.


Experiment series 1

Protein A

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta

Protein B

Protein: BRAF
Fragment: 334-343
Site: 14-3-3_pSer339
Modification: (SEP)339
Sequence: PSPSKSIPIP
Construct: PSPSK(pSer)IPIP (crude synthetic peptide)
Average holdup BI: 0.33
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 132
P value: 4.48
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.58
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: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta

Protein B

Protein: BRAF
Fragment: 360-369
Site: 14-3-3_pSer365
Modification: (SEP)365
Sequence: RDRSSSAPNV
Construct: RDRSS(pSer)APNV (crude synthetic peptide)
Measured dissociation constant (10-6M): <10
Immobilized partner concentration (10-6M): 132
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 1
Measured pKd: 5
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: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta

Protein B

Protein: BRAF
Fragment: 389-398
Site: 14-3-3_pSer394
Modification: (SEP)394
Sequence: RGDGGSTTGL
Construct: RGDGG(pSer)TTGL (crude synthetic peptide)
Average holdup BI: -0.04
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 132
P value: 0.75
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 4

Protein A

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta

Protein B

Protein: BRAF
Fragment: 441-450
Site: 14-3-3_pSer446
Modification: (SEP)446
Sequence: LGRRDSSDDW
Construct: LGRRD(pSer)SDDW (crude synthetic peptide)
Average holdup BI: -0.05
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 132
P value: 0.7
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: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta

Protein B

Protein: BRAF
Fragment: 462-471
Site: 14-3-3_pSer467
Modification: (SEP)467
Sequence: RIGSGSFGTV
Construct: RIGSG(pSer)FGTV (crude synthetic peptide)
Average holdup BI: 0.14
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 132
P value: 3.25
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: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta

Protein B

Protein: BRAF
Fragment: 574-583
Site: 14-3-3_pSer579
Modification: (SEP)579
Sequence: HRDLKSNNIF
Construct: HRDLK(pSer)NNIF (crude synthetic peptide)
Average holdup BI: 0.14
Holdup BI standard deviation: 0.39
Immobilized partner concentration (10-6M): 132
P value: 0.31
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: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta

Protein B

Protein: BRAF
Fragment: 597-606
Site: 14-3-3_pSer602
Modification: (SEP)602
Sequence: LATVKSRWSG
Construct: LATVK(pSer)RWSG (crude synthetic peptide)
Average holdup BI: 0
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 132
P value: 0.03
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: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta

Protein B

Protein: BRAF
Fragment: 609-618
Site: 14-3-3_pSer614
Modification: (SEP)614
Sequence: QFEQLSGSIL
Construct: QFEQL(pSer)GSIL (crude synthetic peptide)
Average holdup BI: -0.25
Holdup BI standard deviation: 0.31
Immobilized partner concentration (10-6M): 132
P value: 1.1
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 9

Protein A

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta

Protein B

Protein: BRAF
Fragment: 724-733
Site: 14-3-3_pSer729
Modification: (SEP)729
Sequence: IHRSASEPSL
Construct: IHRSA(pSer)EPSL (crude synthetic peptide)
Average holdup BI: 0.95
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 132
P value: 6.06
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 5.17
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: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta

Protein B

Protein: BRAF
Fragment: 465-474
Site: 14-3-3_pThr470
Modification: (TPO)470
Sequence: SGSFGTVYKG
Construct: SGSFG(pThr)VYKG (crude synthetic peptide)
Average holdup BI: 0.34
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 132
P value: 6.01
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.59
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: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta

Protein B

Protein: BRAF
Fragment: 594-603
Site: 14-3-3_pThr599
Modification: (TPO)599
Sequence: DFGLATVKSR
Construct: DFGLA(pThr)VKSR (crude synthetic peptide)
Average holdup BI: 0.03
Holdup BI standard deviation: 0.08
Immobilized partner concentration (10-6M): 132
P value: 0.38
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))