Interaction Details


Details of experiments between O60825 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: PFKFB2
Fragment: 461-470
Site: 14-3-3_pSer466
Modification: (SEP)466
Sequence: RMRRNSFTPL
Construct: RMRRN(pSer)FTPL (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: 0.38
Holdup BI standard deviation: 0.15
Immobilized partner concentration (10-6M): 143
P value: 2.49
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.63
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: PFKFB2
Fragment: 478-487
Site: 14-3-3_pSer483
Modification: (SEP)483
Sequence: RPRNYSVGSR
Construct: RPRNY(pSer)VGSR (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: 0.16
Holdup BI standard deviation: 0.11
Immobilized partner concentration (10-6M): 116
P value: 1.33
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 3

Protein A

Protein: PFKFB2
Fragment: 478-487
Site: 14-3-3_pSer483
Modification: (SEP)483
Sequence: RPRNYSVGSR
Construct: RPRNY(pSer)VGSR (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: -0.02
Holdup BI standard deviation: 0.39
Immobilized partner concentration (10-6M): 143
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 4

Protein A

Protein: PFKFB2
Fragment: 470-479
Site: 14-3-3_pThr475
Modification: (TPO)475
Sequence: LSSSNTIRRP
Construct: LSSSN(pThr)IRRP (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: 0.58
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 116
P value: 5.19
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.07
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: PFKFB2
Fragment: 470-479
Site: 14-3-3_pThr475
Modification: (TPO)475
Sequence: LSSSNTIRRP
Construct: LSSSN(pThr)IRRP (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: 0.51
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 143
P value: 5.56
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.86
DOI: 10.1101/2022.12.08.519103
Experimental Details

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