Interaction Details


Details of experiments between Q8IVT5 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: KSR1
Fragment: 326-335
Site: 14-3-3_pSer331
Modification: (SEP)331
Sequence: MRFDLSHGSP
Construct: MRFDL(pSer)HGSP (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: -0.13
Holdup BI standard deviation: 0.33
Immobilized partner concentration (10-6M): 116
P value: 0.34
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: KSR1
Fragment: 326-335
Site: 14-3-3_pSer331
Modification: (SEP)331
Sequence: MRFDLSHGSP
Construct: MRFDL(pSer)HGSP (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: 0.15
Immobilized partner concentration (10-6M): 143
P value: 0.39
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 3

Protein A

Protein: KSR1
Fragment: 401-410
Site: 14-3-3_pSer406
Modification: (SEP)406
Sequence: LRRTESVPSD
Construct: LRRTE(pSer)VPSD (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: 0.25
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 143
P value: 4.82
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.37
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: KSR1
Fragment: 401-410
Site: 14-3-3_pSer406
Modification: (SEP)406
Sequence: LRRTESVPSD
Construct: LRRTE(pSer)VPSD (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: 0.18
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 116
P value: 3.05
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: KSR1
Fragment: 268-277
Site: 14-3-3_pThr273
Modification: (TPO)273
Sequence: FITPPTTPQL
Construct: FITPP(pThr)TPQL (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: -0.26
Holdup BI standard deviation: 0.26
Immobilized partner concentration (10-6M): 116
P value: 1.18
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 6

Protein A

Protein: KSR1
Fragment: 268-277
Site: 14-3-3_pThr273
Modification: (TPO)273
Sequence: FITPPTTPQL
Construct: FITPP(pThr)TPQL (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: -0.32
Holdup BI standard deviation: 0.26
Immobilized partner concentration (10-6M): 143
P value: 1.97
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))