Interaction Details


Details of experiments between Q32P44 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: EML3
Fragment: 171-180
Site: 14-3-3_pSer176
Modification: (SEP)176
Sequence: SRKAISSANL
Construct: SRKAI(pSer)SANL (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: 0.39
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 143
P value: 6.42
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.66
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: EML3
Fragment: 171-180
Site: 14-3-3_pSer176
Modification: (SEP)176
Sequence: SRKAISSANL
Construct: SRKAI(pSer)SANL (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: 0.33
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 116
P value: 7.11
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.62
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: EML3
Fragment: 192-201
Site: 14-3-3_pSer197
Modification: (SEP)197
Sequence: GKDPLSSPGG
Construct: GKDPL(pSer)SPGG (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: 0.15
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 143
P value: 6.17
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.08
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 4

Protein A

Protein: EML3
Fragment: 192-201
Site: 14-3-3_pSer197
Modification: (SEP)197
Sequence: GKDPLSSPGG
Construct: GKDPL(pSer)SPGG (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: 3.53
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: EML3
Fragment: 202-211
Site: 14-3-3_pSer207
Modification: (SEP)207
Sequence: PGSRRSNYNL
Construct: PGSRR(pSer)NYNL (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: 0.05
Immobilized partner concentration (10-6M): 116
P value: 0.89
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 6

Protein A

Protein: EML3
Fragment: 202-211
Site: 14-3-3_pSer207
Modification: (SEP)207
Sequence: PGSRRSNYNL
Construct: PGSRR(pSer)NYNL (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): 143
P value: 0.38
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))