Interaction Details


Details of experiments between Q9P0L2 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: MARK1
Fragment: 644-653
Site: 14-3-3_pSer649
Modification: (SEP)649
Sequence: ARRGTSTGII
Construct: ARRGT(pSer)TGII (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: 0.31
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 116
P value: 6.65
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.59
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: MARK1
Fragment: 644-653
Site: 14-3-3_pSer649
Modification: (SEP)649
Sequence: ARRGTSTGII
Construct: ARRGT(pSer)TGII (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: 0.35
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 143
P value: 4.74
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 3

Protein A

Protein: MARK1
Fragment: 60-69
Site: 14-3-3_pThr65
Modification: (TPO)65
Sequence: YRLQKTIGKG
Construct: YRLQK(pThr)IGKG (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: 0.03
Holdup BI standard deviation: 0.11
Immobilized partner concentration (10-6M): 116
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 4

Protein A

Protein: MARK1
Fragment: 60-69
Site: 14-3-3_pThr65
Modification: (TPO)65
Sequence: YRLQKTIGKG
Construct: YRLQK(pThr)IGKG (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: 0.07
Holdup BI standard deviation: 0.1
Immobilized partner concentration (10-6M): 143
P value: 0.8
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))