Interaction Details


Details of experiments between P31947 and Q14153
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: FAM53B
Fragment: 164-173
Site: 14-3-3_pSer169
Modification: (SEP)169
Sequence: RSSSFSLPSR
Construct: RSSSF(pSer)LPSR (crude synthetic peptide)
Average holdup BI: 0.96
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 113
P value: 7.84
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 5.29
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: FAM53B
Fragment: 164-173
Site: 14-3-3_pSer169
Modification: (SEP)169
Sequence: RSSSFSLPSR
Construct: RSSSF(pSer)LPSR (crude synthetic peptide)
Average holdup BI: 0.95
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 142
P value: 7.73
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 5.17
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
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: FAM53B
Fragment: 164-173
Site: 14-3-3_pSer169
Modification: (SEP)169
Sequence: RSSSFSLPSR
Construct: biotin-ttds-RSSSF(pSer)LPSR (HPLC-purified synthetic peptide)
Measured dissociation constant (10-6M): 15
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: 2022-02-09
Number of measurements: 3
Measured pKd: 4.82
DOI: 10.1101/2022.12.08.519103

Experiment series 4

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: FAM53B
Fragment: 164-173
Site: 14-3-3_pSer169
Modification: (SEP)169
Sequence: RSSSFSLPSR
Construct: RSSSF(pSer)LPSR (crude synthetic peptide)
Average holdup BI: 0.95
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 111
P value: 7.09
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 5.25
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

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: FAM53B
Fragment: 263-272
Site: 14-3-3_pSer268
Modification: (SEP)268
Sequence: LSRSRSQPCV
Construct: LSRSR(pSer)QPCV (crude synthetic peptide)
Average holdup BI: -0.31
Holdup BI standard deviation: 0.61
Immobilized partner concentration (10-6M): 142
P value: 0.64
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: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: FAM53B
Fragment: 263-272
Site: 14-3-3_pSer268
Modification: (SEP)268
Sequence: LSRSRSQPCV
Construct: LSRSR(pSer)QPCV (crude synthetic peptide)
Average holdup BI: -0.08
Holdup BI standard deviation: 0.78
Immobilized partner concentration (10-6M): 111
P value: 0.07
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))