Interaction Details


Details of experiments between P33176 and Q04917
Note that the results of all experiments are listed, regardless of the modification states of the fragments.



Experiment series 1

Protein A

Protein: KIF5B
Fragment: 149-158
Site: 14-3-3_pSer154
Modification: (SEP)154
Sequence: SKTNLSVHED
Construct: SKTNL(pSer)VHED (crude synthetic peptide)

Protein B

Protein: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta
Average holdup BI: 0.01
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 130
P value: 0.23
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 2

Protein A

Protein: KIF5B
Fragment: 933-942
Site: 14-3-3_pSer938
Modification: (SEP)938
Sequence: SPTHPSAIRG
Construct: SPTHP(pSer)AIRG (crude synthetic peptide)

Protein B

Protein: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta
Average holdup BI: 0
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 130
P value: 0.09
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 3

Protein A

Protein: KIF5B
Fragment: 945-954
Site: 14-3-3_pSer950
Modification: (SEP)950
Sequence: AFVQNSQPVA
Construct: AFVQN(pSer)QPVA (crude synthetic peptide)

Protein B

Protein: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta
Average holdup BI: 0.13
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 130
P value: 2.69
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 4

Protein A

Protein: KIF5B
Fragment: 392-401
Site: 14-3-3_pThr397
Modification: (TPO)397
Sequence: VDKDITLTND
Construct: VDKDI(pThr)LTND (crude synthetic peptide)

Protein B

Protein: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta
Average holdup BI: 0.01
Holdup BI standard deviation: 0.1
Immobilized partner concentration (10-6M): 130
P value: 0.1
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 5

Protein A

Protein: KIF5B
Fragment: 446-455
Site: 14-3-3_pThr451
Modification: (TPO)451
Sequence: VEKLKTQMLD
Construct: VEKLK(pThr)QMLD (crude synthetic peptide)

Protein B

Protein: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta
Average holdup BI: 0.44
Holdup BI standard deviation: 0.35
Immobilized partner concentration (10-6M): 130
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
Measured pKd: 3.78
DOI: 10.1101/2022.12.08.519103
Experimental Details

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