Interaction Details


Details of experiments between Q6ICG6 and P62258
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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.08
Holdup BI standard deviation: 0.08
Immobilized partner concentration (10-6M): 116
P value: 0.99
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 2

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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.01
Holdup BI standard deviation: 0.12
Immobilized partner concentration (10-6M): 143
P value: 0.07
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: KIAA0930
Fragment: 284-293
Site: 14-3-3_pSer289
Modification: (SEP)289
Sequence: RVTSFSTPPT
Construct: RVTSF(pSer)TPPT (crude synthetic peptide)

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.8
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 116
P value: 9.85
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.54
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: KIAA0930
Fragment: 284-293
Site: 14-3-3_pSer289
Modification: (SEP)289
Sequence: RVTSFSTPPT
Construct: RVTSF(pSer)TPPT (crude synthetic peptide)

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.81
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 143
P value: 9.1
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.48
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: 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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Measured dissociation constant (10-6M): 42
Standard deviation of dissociation constant (10-6M): 14
Experiment method: competitive Fluorescence Polarization
Details of affinity fitting: Competitive binding equation in ProFit
Date of measurement: 2022-02-09
Number of measurements: 3
Measured pKd: 4.38
DOI: 10.1101/2022.12.08.519103

Experiment series 6

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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: -0.07
Holdup BI standard deviation: 0.24
Immobilized partner concentration (10-6M): 116
P value: 0.26
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: KIAA0930
Fragment: 301-310
Site: 14-3-3_pSer306
Modification: (SEP)306
Sequence: AFFSPSLKRK
Construct: AFFSP(pSer)LKRK (crude synthetic peptide)

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0
Holdup BI standard deviation: 0.12
Immobilized partner concentration (10-6M): 143
P value: 0.01
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: KIAA0930
Fragment: 357-366
Site: 14-3-3_pSer362
Modification: (SEP)362
Sequence: RSLVGSWLKL
Construct: RSLVG(pSer)WLKL (crude synthetic peptide)

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.07
Holdup BI standard deviation: 0.89
Immobilized partner concentration (10-6M): 116
P value: 0.05
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 9

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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: -0.14
Holdup BI standard deviation: 0.33
Immobilized partner concentration (10-6M): 116
P value: 0.24
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 10

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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: -0.07
Holdup BI standard deviation: 0.25
Immobilized partner concentration (10-6M): 143
P value: 0.24
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))