Interaction Details


Details of experiments between P31947 and Q8IVT5
Note that the results of all experiments are listed, regardless of the modification states of the fragments.


Experiment series 1

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: KSR1
Fragment: 306-315
Site: 14-3-3_pSer311
Modification: (SEP)311
Sequence: LTRSKSHESQ
Construct: LTRSK(pSer)HESQ (crude synthetic peptide)
Average holdup BI: 0.15
Holdup BI standard deviation: 0.5
Immobilized partner concentration (10-6M): 113
P value: 0.12
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 2

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: KSR1
Fragment: 326-335
Site: 14-3-3_pSer331
Modification: (SEP)331
Sequence: MRFDLSHGSP
Construct: MRFDL(pSer)HGSP (crude synthetic peptide)
Average holdup BI: -0.14
Holdup BI standard deviation: 0.2
Immobilized partner concentration (10-6M): 113
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 3

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: KSR1
Fragment: 326-335
Site: 14-3-3_pSer331
Modification: (SEP)331
Sequence: MRFDLSHGSP
Construct: MRFDL(pSer)HGSP (crude synthetic peptide)
Average holdup BI: -0.06
Holdup BI standard deviation: 0.15
Immobilized partner concentration (10-6M): 142
P value: 0.43
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: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: KSR1
Fragment: 326-335
Site: 14-3-3_pSer331
Modification: (SEP)331
Sequence: MRFDLSHGSP
Construct: MRFDL(pSer)HGSP (crude synthetic peptide)
Average holdup BI: -0.15
Holdup BI standard deviation: 0.21
Immobilized partner concentration (10-6M): 111
P value: 0.68
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: KSR1
Fragment: 401-410
Site: 14-3-3_pSer406
Modification: (SEP)406
Sequence: LRRTESVPSD
Construct: LRRTE(pSer)VPSD (crude synthetic peptide)
Average holdup BI: 0.12
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 113
P value: 2.23
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 6

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: KSR1
Fragment: 401-410
Site: 14-3-3_pSer406
Modification: (SEP)406
Sequence: LRRTESVPSD
Construct: LRRTE(pSer)VPSD (crude synthetic peptide)
Average holdup BI: 0.19
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 142
P value: 4.56
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: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: KSR1
Fragment: 401-410
Site: 14-3-3_pSer406
Modification: (SEP)406
Sequence: LRRTESVPSD
Construct: LRRTE(pSer)VPSD (crude synthetic peptide)
Average holdup BI: 0.28
Holdup BI standard deviation: 0.14
Immobilized partner concentration (10-6M): 111
P value: 1.99
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.54
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: KSR1
Fragment: 268-277
Site: 14-3-3_pThr273
Modification: (TPO)273
Sequence: FITPPTTPQL
Construct: FITPP(pThr)TPQL (crude synthetic peptide)
Average holdup BI: 0
Holdup BI standard deviation: 0.28
Immobilized partner concentration (10-6M): 113
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 9

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: KSR1
Fragment: 268-277
Site: 14-3-3_pThr273
Modification: (TPO)273
Sequence: FITPPTTPQL
Construct: FITPP(pThr)TPQL (crude synthetic peptide)
Average holdup BI: -0.38
Holdup BI standard deviation: 0.27
Immobilized partner concentration (10-6M): 142
P value: 2.41
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 10

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: KSR1
Fragment: 268-277
Site: 14-3-3_pThr273
Modification: (TPO)273
Sequence: FITPPTTPQL
Construct: FITPP(pThr)TPQL (crude synthetic peptide)
Average holdup BI: 0.01
Holdup BI standard deviation: 0.1
Immobilized partner concentration (10-6M): 111
P value: 0.08
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))