Interaction Details


Details of experiments between P62258 and Q6WCQ1
Note that the results of all experiments are listed, regardless of the modification states of the fragments.



Experiment series 1

Protein A

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Protein B

Protein: MPRIP
Fragment: 1011-1020
Site: 14-3-3_pSer1016
Modification: (SEP)1016
Sequence: NIRSKSVIEQ
Construct: NIRSK(pSer)VIEQ (crude synthetic peptide)
Average holdup BI: 0.27
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 143
P value: 5.05
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.42
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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Protein B

Protein: MPRIP
Fragment: 1011-1020
Site: 14-3-3_pSer1016
Modification: (SEP)1016
Sequence: NIRSKSLKEG
Construct: NIRSK(pSer)LKEG (crude synthetic peptide)
Average holdup BI: 0.06
Holdup BI standard deviation: 0.92
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 3

Protein A

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Protein B

Protein: MPRIP
Fragment: 1011-1020
Site: 14-3-3_pSer1016
Modification: (SEP)1016
Sequence: NIRSKSVIEQ
Construct: NIRSK(pSer)VIEQ (crude synthetic peptide)
Average holdup BI: 0.25
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 116
P value: 3.78
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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Protein B

Protein: MPRIP
Fragment: 264-273
Site: 14-3-3_pSer269
Modification: (SEP)269
Sequence: ESGYFSLEKT
Construct: ESGYF(pSer)LEKT (crude synthetic peptide)
Average holdup BI: 0.06
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 116
P value: 1.52
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 5

Protein A

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Protein B

Protein: MPRIP
Fragment: 264-273
Site: 14-3-3_pSer269
Modification: (SEP)269
Sequence: ESGYFSLEKT
Construct: ESGYF(pSer)LEKT (crude synthetic peptide)
Average holdup BI: 0.1
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 143
P value: 2.03
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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Protein B

Protein: MPRIP
Fragment: 289-298
Site: 14-3-3_pSer294
Modification: (SEP)294
Sequence: SPPSPSTPNH
Construct: SPPSP(pSer)TPNH (crude synthetic peptide)
Average holdup BI: 0.03
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 116
P value: 0.36
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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Protein B

Protein: MPRIP
Fragment: 289-298
Site: 14-3-3_pSer294
Modification: (SEP)294
Sequence: SPPSPSTPNH
Construct: SPPSP(pSer)TPNH (crude synthetic peptide)
Average holdup BI: 0.09
Holdup BI standard deviation: 0.08
Immobilized partner concentration (10-6M): 143
P value: 1.21
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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Protein B

Protein: MPRIP
Fragment: 296-305
Site: 14-3-3_pSer301
Modification: (SEP)301
Sequence: PNHRRSQVIE
Construct: PNHRR(pSer)QVIE (crude synthetic peptide)
Average holdup BI: -0.09
Holdup BI standard deviation: 0.48
Immobilized partner concentration (10-6M): 116
P value: 0.09
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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Protein B

Protein: MPRIP
Fragment: 988-997
Site: 14-3-3_pSer993
Modification: (SEP)993
Sequence: IMKSKSNPDF
Construct: IMKSK(pSer)NPDF (crude synthetic peptide)
Average holdup BI: 0.98
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 116
P value: 6.35
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 5.6
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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Protein B

Protein: MPRIP
Fragment: 988-997
Site: 14-3-3_pSer993
Modification: (SEP)993
Sequence: IMKSKSNPDF
Construct: IMKSK(pSer)NPDF (crude synthetic peptide)
Average holdup BI: 0.98
Holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 143
P value: 10.05
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 5.53
DOI: 10.1101/2022.12.08.519103
Experimental Details

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


Experiment series 11

Protein A

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Protein B

Protein: MPRIP
Fragment: 373-382
Site: 14-3-3_pThr378
Modification: (TPO)378
Sequence: LDRRSTEPSV
Construct: LDRRS(pThr)EPSV (crude synthetic peptide)
Average holdup BI: 0.59
Holdup BI standard deviation: 0.13
Immobilized partner concentration (10-6M): 116
P value: 3.81
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.1
DOI: 10.1101/2022.12.08.519103
Experimental Details

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


Experiment series 12

Protein A

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Protein B

Protein: MPRIP
Fragment: 373-382
Site: 14-3-3_pThr378
Modification: (TPO)378
Sequence: LDRRSTEPSV
Construct: LDRRS(pThr)EPSV (crude synthetic peptide)
Average holdup BI: 0.55
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 143
P value: 7.06
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.94
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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Protein B

Protein: MPRIP
Fragment: 537-546
Site: 14-3-3_pThr542
Modification: (TPO)542
Sequence: EGRSKTFDWA
Construct: EGRSK(pThr)FDWA (crude synthetic peptide)
Average holdup BI: 0.54
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 116
P value: 6.06
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.01
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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Protein B

Protein: MPRIP
Fragment: 537-546
Site: 14-3-3_pThr542
Modification: (TPO)542
Sequence: EGRSKTFDWA
Construct: EGRSK(pThr)FDWA (crude synthetic peptide)
Average holdup BI: 0.53
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 143
P value: 5.72
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))