Interaction Details


Details of experiments between P27448 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: MARK3
Fragment: 535-544
Site: 14-3-3_pSer540
Modification: (SEP)540
Sequence: RTPVASTHSI
Construct: RTPVA(pSer)THSI (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: 0.06
Immobilized partner concentration (10-6M): 116
P value: 0.18
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 2

Protein A

Protein: MARK3
Fragment: 535-544
Site: 14-3-3_pSer540
Modification: (SEP)540
Sequence: RTPVASTHSI
Construct: RTPVA(pSer)THSI (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: 0.1
Immobilized partner concentration (10-6M): 143
P value: 0.28
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: MARK3
Fragment: 538-547
Site: 14-3-3_pSer543
Modification: (SEP)543
Sequence: VASTHSISSA
Construct: VASTH(pSer)ISSA (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: 0.06
Immobilized partner concentration (10-6M): 116
P value: 0.19
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: MARK3
Fragment: 538-547
Site: 14-3-3_pSer543
Modification: (SEP)543
Sequence: VASTHSISSA
Construct: VASTH(pSer)ISSA (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: 0.06
Immobilized partner concentration (10-6M): 143
P value: 0.63
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 5

Protein A

Protein: MARK3
Fragment: 596-605
Site: 14-3-3_pSer601
Modification: (SEP)601
Sequence: TRSRGSTNLF
Construct: TRSRG(pSer)TNLF (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: 0.17
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 143
P value: 3.23
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.15
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: MARK3
Fragment: 596-605
Site: 14-3-3_pSer601
Modification: (SEP)601
Sequence: TRSRGSTNLF
Construct: TRSRG(pSer)TNLF (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: 0.14
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 116
P value: 4.15
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: MARK3
Fragment: 91-100
Site: 14-3-3_pSer96
Modification: (SEP)96
Sequence: QLNPTSLQKL
Construct: QLNPT(pSer)LQKL (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: 0.07
Immobilized partner concentration (10-6M): 116
P value: 0.3
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 8

Protein A

Protein: MARK3
Fragment: 91-100
Site: 14-3-3_pSer96
Modification: (SEP)96
Sequence: QLNPTSLQKL
Construct: QLNPT(pSer)LQKL (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: 0.11
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 9

Protein A

Protein: MARK3
Fragment: 206-215
Site: 14-3-3_pThr211
Modification: (TPO)211
Sequence: GGKLDTFCGS
Construct: GGKLD(pThr)FCGS (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: 1.24
Immobilized partner concentration (10-6M): 116
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 10

Protein A

Protein: MARK3
Fragment: 486-495
Site: 14-3-3_pThr491
Modification: (TPO)491
Sequence: RKKSSTVPSS
Construct: RKKSS(pThr)VPSS (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: 0.52
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: MARK3
Fragment: 486-495
Site: 14-3-3_pThr491
Modification: (TPO)491
Sequence: RKKSSTVPSS
Construct: RKKSS(pThr)VPSS (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: 0.05
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 143
P value: 0.73
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: MARK3
Fragment: 85-94
Site: 14-3-3_pThr90
Modification: (TPO)90
Sequence: KIIDKTQLNP
Construct: KIIDK(pThr)QLNP (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: 0.09
Immobilized partner concentration (10-6M): 116
P value: 1.8
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 13

Protein A

Protein: MARK3
Fragment: 85-94
Site: 14-3-3_pThr90
Modification: (TPO)90
Sequence: KIIDKTQLNP
Construct: KIIDK(pThr)QLNP (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: -0.08
Holdup BI standard deviation: 0.1
Immobilized partner concentration (10-6M): 143
P value: 0.93
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))