Interaction Details


Details of experiments between P62258 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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

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: 0.92
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 116
P value: 8
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 5.02
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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

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: 0.91
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 143
P value: 7.59
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.84
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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

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): 24
Standard deviation of dissociation constant (10-6M): 4
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: 4.62
DOI: 10.1101/2022.12.08.519103

Experiment series 4

Protein A

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

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: -0.13
Holdup BI standard deviation: 0.49
Immobilized partner concentration (10-6M): 143
P value: 0.2
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))