Interaction Details


Details of experiments between P62258 and P56524
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: HDAC4
Fragment: 260-269
Site: 14-3-3_pSer265
Modification: (SEP)265
Sequence: VAERRSSPLL
Construct: VAERR(pSer)SPLL (crude synthetic peptide)
Average holdup BI: 0.78
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 143
P value: 7.66
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.38
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: HDAC4
Fragment: 260-269
Site: 14-3-3_pSer265
Modification: (SEP)265
Sequence: VAERRSSPLL
Construct: VAERR(pSer)SPLL (crude synthetic peptide)
Average holdup BI: 0.72
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 116
P value: 8.26
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.34
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: HDAC4
Fragment: 345-354
Site: 14-3-3_pSer350
Modification: (SEP)350
Sequence: LYTSPSLPNI
Construct: LYTSP(pSer)LPNI (crude synthetic peptide)
Average holdup BI: 0.61
Holdup BI standard deviation: 0.11
Immobilized partner concentration (10-6M): 116
P value: 3.33
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.13
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: HDAC4
Fragment: 345-354
Site: 14-3-3_pSer350
Modification: (SEP)350
Sequence: LYTSPSLPNI
Construct: LYTSP(pSer)LPNI (crude synthetic peptide)
Average holdup BI: 0.56
Holdup BI standard deviation: 0.15
Immobilized partner concentration (10-6M): 143
P value: 2.29
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 5

Protein A

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

Protein B

Protein: HDAC4
Fragment: 579-588
Site: 14-3-3_pSer584
Modification: (SEP)584
Sequence: GQRQPSEQEL
Construct: GQRQP(pSer)EQEL (crude synthetic peptide)
Average holdup BI: -0.02
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 116
P value: 1.21
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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Protein B

Protein: HDAC4
Fragment: 579-588
Site: 14-3-3_pSer584
Modification: (SEP)584
Sequence: GQRQPSEQEL
Construct: GQRQP(pSer)EQEL (crude synthetic peptide)
Average holdup BI: -0.01
Holdup BI standard deviation: 0.03
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))

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: HDAC4
Fragment: 627-636
Site: 14-3-3_pSer632
Modification: (SEP)632
Sequence: LSRAQSSPAS
Construct: LSRAQ(pSer)SPAS (crude synthetic peptide)
Average holdup BI: 0.91
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 116
P value: 3.45
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.96
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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Protein B

Protein: HDAC4
Fragment: 627-636
Site: 14-3-3_pSer632
Modification: (SEP)632
Sequence: LSRAQSSPAS
Construct: LSRAQ(pSer)SPAS (crude synthetic peptide)
Average holdup BI: 0.89
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 143
P value: 4.23
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.74
DOI: 10.1101/2022.12.08.519103
Experimental Details

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