Interaction Details


Details of experiments between Q6ICG6 and P63104
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: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta
Average holdup BI: 0.06
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 132
P value: 1.71
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: RVTSF(pSer)TPPT (crude synthetic peptide)

Protein B

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta
Average holdup BI: 0.92
Holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 132
P value: 9.43
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.91
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: 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: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta
Measured dissociation constant (10-6M): 21
Standard deviation of dissociation constant (10-6M): 2
Experiment method: competitive Fluorescence Polarization
Details of affinity fitting: Competitive binding equation in ProFit
Date of measurement: 2022-02-10
Number of measurements: 3
Measured pKd: 4.69
DOI: 10.1101/2022.12.08.519103

Experiment series 4

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: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta
Measured dissociation constant (10-6M): 25
Standard deviation of dissociation constant (10-6M): 2
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: 4.59
DOI: 10.1101/2022.12.08.519103

Experiment series 5

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: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta
Average holdup BI: 0.12
Holdup BI standard deviation: 0.17
Immobilized partner concentration (10-6M): 132
P value: 0.66
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 6

Protein A

Protein: KIAA0930
Fragment: 399-408
Site: 14-3-3_pThr404
Modification: (TPO)404
Sequence: KPILMT----
Construct: EVRQKPILM(pThr) (crude synthetic peptide)

Protein B

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta
Average holdup BI: 0.05
Holdup BI standard deviation: 0.23
Immobilized partner concentration (10-6M): 132
P value: 0.17
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))