Interaction Details


Details of experiments between P31947 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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: 0.62
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 142
P value: 6.61
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.05
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: 0.55
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 113
P value: 7.19
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.04
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: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: 0.48
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 111
P value: 6.4
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.92
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: 0.58
Holdup BI standard deviation: 0.12
Immobilized partner concentration (10-6M): 113
P value: 3.06
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.08
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: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: 0.49
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 111
P value: 5.93
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.93
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: 0.04
Immobilized partner concentration (10-6M): 113
P value: 0.22
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: 0.03
Immobilized partner concentration (10-6M): 142
P value: 0.67
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 8

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: -0.04
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 111
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 9

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: 0.94
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 142
P value: 4.41
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 5
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: 0.92
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 113
P value: 3.48
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.99
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: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: 0.86
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 111
P value: 2.25
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.75
DOI: 10.1101/2022.12.08.519103
Experimental Details

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