Interaction Details


Details of experiments between P31947 and Q6ICG6
Note that the results of all experiments are listed, regardless of the modification states of the fragments.


Experiment series 1

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: KIAA0930
Fragment: 257-266
Site: 14-3-3_pSer262
Modification: (SEP)262
Sequence: AVSRVSTGDT
Construct: AVSRV(pSer)TGDT (crude synthetic peptide)
Average holdup BI: 0.04
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 113
P value: 1.15
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: KIAA0930
Fragment: 257-266
Site: 14-3-3_pSer262
Modification: (SEP)262
Sequence: AVSRVSTGDT
Construct: AVSRV(pSer)TGDT (crude synthetic peptide)
Average holdup BI: 0.02
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 142
P value: 0.46
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: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: KIAA0930
Fragment: 257-266
Site: 14-3-3_pSer262
Modification: (SEP)262
Sequence: AVSRVSTGDT
Construct: AVSRV(pSer)TGDT (crude synthetic peptide)
Average holdup BI: 0.14
Holdup BI standard deviation: 0.12
Immobilized partner concentration (10-6M): 111
P value: 1.15
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 4

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: KIAA0930
Fragment: 284-293
Site: 14-3-3_pSer289
Modification: (SEP)289
Sequence: RVTSFSTPPT
Construct: RVTSF(pSer)TPPT (crude synthetic peptide)
Average holdup BI: 0.87
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 142
P value: 9.08
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.67
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: KIAA0930
Fragment: 284-293
Site: 14-3-3_pSer289
Modification: (SEP)289
Sequence: RVTSFSTPPT
Construct: RVTSF(pSer)TPPT (crude synthetic peptide)
Average holdup BI: 0.83
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 113
P value: 9.75
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.65
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

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)
Measured dissociation constant (10-6M): 42
Standard deviation of dissociation constant (10-6M): 18
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 7

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: KIAA0930
Fragment: 284-293
Site: 14-3-3_pSer289
Modification: (SEP)289
Sequence: RVTSFSTPPT
Construct: RVTSF(pSer)TPPT (crude synthetic peptide)
Average holdup BI: 0.87
Holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 111
P value: 8.77
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.79
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: KIAA0930
Fragment: 301-310
Site: 14-3-3_pSer306
Modification: (SEP)306
Sequence: AFFSPSLKRK
Construct: AFFSP(pSer)LKRK (crude synthetic peptide)
Average holdup BI: -0.12
Holdup BI standard deviation: 0.18
Immobilized partner concentration (10-6M): 113
P value: 0.63
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: KIAA0930
Fragment: 301-310
Site: 14-3-3_pSer306
Modification: (SEP)306
Sequence: AFFSPSLKRK
Construct: AFFSP(pSer)LKRK (crude synthetic peptide)
Average holdup BI: -0.09
Holdup BI standard deviation: 0.27
Immobilized partner concentration (10-6M): 142
P value: 0.28
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 10

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: KIAA0930
Fragment: 301-310
Site: 14-3-3_pSer306
Modification: (SEP)306
Sequence: AFFSPSLKRK
Construct: AFFSP(pSer)LKRK (crude synthetic peptide)
Average holdup BI: 0.13
Holdup BI standard deviation: 0.21
Immobilized partner concentration (10-6M): 111
P value: 0.55
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 11

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: KIAA0930
Fragment: 357-366
Site: 14-3-3_pSer362
Modification: (SEP)362
Sequence: RSLVGSWLKL
Construct: RSLVG(pSer)WLKL (crude synthetic peptide)
Average holdup BI: -0.31
Holdup BI standard deviation: 0.95
Immobilized partner concentration (10-6M): 113
P value: 0.3
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 12

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: KIAA0930
Fragment: 399-408
Site: 14-3-3_pThr404
Modification: (TPO)404
Sequence: KPILMT----
Construct: EVRQKPILM(pThr) (crude synthetic peptide)
Average holdup BI: -0.22
Holdup BI standard deviation: 0.24
Immobilized partner concentration (10-6M): 113
P value: 0.58
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 13

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: KIAA0930
Fragment: 399-408
Site: 14-3-3_pThr404
Modification: (TPO)404
Sequence: KPILMT----
Construct: EVRQKPILM(pThr) (crude synthetic peptide)
Average holdup BI: -0.14
Holdup BI standard deviation: 0.28
Immobilized partner concentration (10-6M): 142
P value: 0.49
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))