Interaction Details


Details of experiments between Q6ICG6 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: KIAA0930
Fragment: 257-266
Site: 14-3-3_pSer262
Modification: (SEP)262
Sequence: AVSRVSTGDT
Construct: AVSRV(pSer)TGDT (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: 0.06
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 130
P value: 1.28
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: KIAA0930
Fragment: 284-293
Site: 14-3-3_pSer289
Modification: (SEP)289
Sequence: RVTSFSTPPT
Construct: biotin-ttds-RVTSF(pSer)TPPT (HPLC-purified synthetic peptide)

Protein B

Protein: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta
Measured dissociation constant (10-6M): 9.5
Standard deviation of dissociation constant (10-6M): 0.9
Experiment method: competitive Fluorescence Polarization
Details of affinity fitting: Competitive binding equation in ProFit
Date of measurement: 2022-01-19
Number of measurements: 3
Measured pKd: 5.02
DOI: 10.1101/2022.12.08.519103

Experiment series 3

Protein A

Protein: KIAA0930
Fragment: 284-293
Site: 14-3-3_pSer289
Modification: (SEP)289
Sequence: RVTSFSTPPT
Construct: RVTSF(pSer)TPPT (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: 0.92
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 130
P value: 8.45
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.93
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: KIAA0930
Fragment: 301-310
Site: 14-3-3_pSer306
Modification: (SEP)306
Sequence: AFFSPSLKRK
Construct: AFFSP(pSer)LKRK (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: 0.27
Holdup BI standard deviation: 0.2
Immobilized partner concentration (10-6M): 130
P value: 1.45
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.46
DOI: 10.1101/2022.12.08.519103
Experimental Details

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