Interaction Details


Details of experiments between P31947 and Q9UQL6
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: HDAC5
Fragment: 254-263
Site: 14-3-3_pSer259
Modification: (SEP)259
Sequence: LRKTASEPNL
Construct: LRKTA(pSer)EPNL (crude synthetic peptide)
Average holdup BI: 0.75
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 142
P value: 9.07
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.32
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: HDAC5
Fragment: 254-263
Site: 14-3-3_pSer259
Modification: (SEP)259
Sequence: LRKTASEPNL
Construct: LRKTA(pSer)EPNL (crude synthetic peptide)
Average holdup BI: 0.7
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 113
P value: 7.72
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.31
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: HDAC5
Fragment: 254-263
Site: 14-3-3_pSer259
Modification: (SEP)259
Sequence: LRKTASEPNL
Construct: LRKTA(pSer)EPNL (crude synthetic peptide)
Average holdup BI: 0.66
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 111
P value: 5.84
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.24
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: HDAC5
Fragment: 493-502
Site: 14-3-3_pSer498
Modification: (SEP)498
Sequence: LSRTQSSPLP
Construct: LSRTQ(pSer)SPLP (crude synthetic peptide)
Average holdup BI: 0.72
Holdup BI standard deviation: 0.11
Immobilized partner concentration (10-6M): 113
P value: 3.56
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.36
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: HDAC5
Fragment: 493-502
Site: 14-3-3_pSer498
Modification: (SEP)498
Sequence: LSRTQSSPLP
Construct: LSRTQ(pSer)SPLP (crude synthetic peptide)
Measured dissociation constant (10-6M): <10
Immobilized partner concentration (10-6M): 111
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 1
Measured pKd: 5
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: HDAC5
Fragment: 542-551
Site: 14-3-3_pThr547
Modification: (TPO)547
Sequence: PRQPTTHPEE
Construct: PRQPT(pThr)HPEE (crude synthetic peptide)
Average holdup BI: 0.05
Holdup BI standard deviation: 0.22
Immobilized partner concentration (10-6M): 113
P value: 0.17
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: HDAC5
Fragment: 542-551
Site: 14-3-3_pThr547
Modification: (TPO)547
Sequence: PRQPTTHPEE
Construct: PRQPT(pThr)HPEE (crude synthetic peptide)
Average holdup BI: 0.18
Holdup BI standard deviation: 0.14
Immobilized partner concentration (10-6M): 142
P value: 1.36
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: HDAC5
Fragment: 542-551
Site: 14-3-3_pThr547
Modification: (TPO)547
Sequence: PRQPTTHPEE
Construct: PRQPT(pThr)HPEE (crude synthetic peptide)
Average holdup BI: -0.14
Holdup BI standard deviation: 0.36
Immobilized partner concentration (10-6M): 111
P value: 0.37
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))