Interaction Details


Details of experiments between P27448 and P61981
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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.11
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 134
P value: 2.14
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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.03
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 129
P value: 1.04
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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 134
P value: 0.03
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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 129
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 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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.5
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 129
P value: 7.52
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.9
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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.5
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 134
P value: 7.16
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 7

Protein A

Protein: MARK3
Fragment: 601-610
Site: 14-3-3_pSer606
Modification: (SEP)606
Sequence: STNLFSKLTS
Construct: STNLF(pSer)KLTS (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: -0.12
Holdup BI standard deviation: 1.22
Immobilized partner concentration (10-6M): 134
P value: 0.07
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 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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.1
Holdup BI standard deviation: 0.09
Immobilized partner concentration (10-6M): 134
P value: 1.1
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 9

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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.2
Holdup BI standard deviation: 0.09
Immobilized partner concentration (10-6M): 129
P value: 2.54
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 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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.09
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 134
P value: 1.81
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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.09
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 129
P value: 1.67
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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0
Holdup BI standard deviation: 0.08
Immobilized partner concentration (10-6M): 134
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 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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: -0.06
Holdup BI standard deviation: 0.1
Immobilized partner concentration (10-6M): 129
P value: 0.64
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 14

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: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.06
Holdup BI standard deviation: 0.09
Immobilized partner concentration (10-6M): 115
P value: 0.61
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 15

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: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: -0.04
Holdup BI standard deviation: 0.08
Immobilized partner concentration (10-6M): 115
P value: 0.44
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 16

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: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.42
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 115
P value: 8.21
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.82
DOI: 10.1101/2022.12.08.519103
Experimental Details

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


Experiment series 17

Protein A

Protein: MARK3
Fragment: 601-610
Site: 14-3-3_pSer606
Modification: (SEP)606
Sequence: STNLFSKLTS
Construct: STNLF(pSer)KLTS (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: -0.04
Holdup BI standard deviation: 0.59
Immobilized partner concentration (10-6M): 115
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 18

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: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.12
Holdup BI standard deviation: 0.09
Immobilized partner concentration (10-6M): 115
P value: 1.37
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 19

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: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.13
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 115
P value: 3.04
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 20

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: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: -0.02
Holdup BI standard deviation: 0.17
Immobilized partner concentration (10-6M): 115
P value: 0.11
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))