Interaction Details


Details of experiments between Q7KZI7 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: MARK2
Fragment: 395-404
Site: 14-3-3_pSer400
Modification: (SEP)400
Sequence: VQRSVSANPK
Construct: VQRSV(pSer)ANPK (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: 0.47
Holdup BI standard deviation: 0.15
Immobilized partner concentration (10-6M): 116
P value: 2.36
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.88
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: MARK2
Fragment: 395-404
Site: 14-3-3_pSer400
Modification: (SEP)400
Sequence: VQRSVSANPK
Construct: VQRSV(pSer)ANPK (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: -0.11
Holdup BI standard deviation: 1.48
Immobilized partner concentration (10-6M): 143
P value: 0.05
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: MARK2
Fragment: 404-413
Site: 14-3-3_pSer409
Modification: (SEP)409
Sequence: KQRRFSDQAA
Construct: KQRRF(pSer)DQAA (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: 0.48
Immobilized partner concentration (10-6M): 116
P value: 0.04
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: MARK2
Fragment: 488-497
Site: 14-3-3_pSer493
Modification: (SEP)493
Sequence: LLERASLGQA
Construct: LLERA(pSer)LGQA (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: 0.05
Immobilized partner concentration (10-6M): 116
P value: 0.57
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: MARK2
Fragment: 488-497
Site: 14-3-3_pSer493
Modification: (SEP)493
Sequence: LLERASLGQA
Construct: LLERA(pSer)LGQA (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
Holdup BI standard deviation: 0.08
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 6

Protein A

Protein: MARK2
Fragment: 566-575
Site: 14-3-3_pSer571
Modification: (SEP)571
Sequence: PVASPSAHNI
Construct: PVASP(pSer)AHNI (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: -0.04
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 116
P value: 1.07
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 7

Protein A

Protein: MARK2
Fragment: 566-575
Site: 14-3-3_pSer571
Modification: (SEP)571
Sequence: PVASPSAHNI
Construct: PVASP(pSer)AHNI (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: -0.01
Holdup BI standard deviation: 0.09
Immobilized partner concentration (10-6M): 143
P value: 0.09
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: MARK2
Fragment: 626-635
Site: 14-3-3_pSer631
Modification: (SEP)631
Sequence: GRRGASGSIF
Construct: GRRGA(pSer)GSIF (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: 0.2
Holdup BI standard deviation: 0.08
Immobilized partner concentration (10-6M): 143
P value: 2.7
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.25
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: MARK2
Fragment: 626-635
Site: 14-3-3_pSer631
Modification: (SEP)631
Sequence: GRRGASGSIF
Construct: GRRGA(pSer)GSIF (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: 0.11
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 116
P value: 3.16
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 10

Protein A

Protein: MARK2
Fragment: 461-470
Site: 14-3-3_pThr466
Modification: (TPO)466
Sequence: LERKKTTPTP
Construct: LERKK(pThr)TPTP (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
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 116
P value: 0.05
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 11

Protein A

Protein: MARK2
Fragment: 461-470
Site: 14-3-3_pThr466
Modification: (TPO)466
Sequence: LERKKTTPTP
Construct: LERKK(pThr)TPTP (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: 0.1
Immobilized partner concentration (10-6M): 143
P value: 0.12
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 12

Protein A

Protein: MARK2
Fragment: 53-62
Site: 14-3-3_pThr58
Modification: (TPO)58
Sequence: YRLLKTIGKG
Construct: YRLLK(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: 0.49
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 143
P value: 5.83
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.83
DOI: 10.1101/2022.12.08.519103
Experimental Details

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


Experiment series 13

Protein A

Protein: MARK2
Fragment: 53-62
Site: 14-3-3_pThr58
Modification: (TPO)58
Sequence: YRLLKTIGKG
Construct: YRLLK(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: 0.3
Holdup BI standard deviation: 0.17
Immobilized partner concentration (10-6M): 116
P value: 1.58
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.57
DOI: 10.1101/2022.12.08.519103
Experimental Details

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


Experiment series 14

Protein A

Protein: MARK2
Fragment: 591-600
Site: 14-3-3_pThr596
Modification: (TPO)596
Sequence: VSSRSTFHAG
Construct: VSSRS(pThr)FHAG (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: 0.72
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 116
P value: 2.28
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.35
DOI: 10.1101/2022.12.08.519103
Experimental Details

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