Interaction Details


Details of experiments between P61981 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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

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: 0.95
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 129
P value: 9.32
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 5.16
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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

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: 0.94
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 134
P value: 8.22
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 5.05
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: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

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: 0.92
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 115
P value: 6.64
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 5.03
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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

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: 0.8
Holdup BI standard deviation: 0.13
Immobilized partner concentration (10-6M): 134
P value: 3.07
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.47
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: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

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): 115
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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

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: 0.32
Holdup BI standard deviation: 0.13
Immobilized partner concentration (10-6M): 129
P value: 2.63
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.55
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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

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: 0.23
Holdup BI standard deviation: 0.13
Immobilized partner concentration (10-6M): 134
P value: 1.65
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 8

Protein A

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

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: 0.11
Holdup BI standard deviation: 0.24
Immobilized partner concentration (10-6M): 115
P value: 0.38
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))