Interaction Details


Details of experiments between Q14153 and P61981
Note that the results of all experiments are listed, regardless of the modification states of the fragments.


Experiment series 1

Protein A

Protein: FAM53B
Fragment: 164-173
Site: 14-3-3_pSer169
Modification: (SEP)169
Sequence: RSSSFSLPSR
Construct: RSSSF(pSer)LPSR (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.99
Holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 134
P value: 6.65
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 5.97
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: FAM53B
Fragment: 164-173
Site: 14-3-3_pSer169
Modification: (SEP)169
Sequence: RSSSFSLPSR
Construct: RSSSF(pSer)LPSR (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.99
Holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 129
P value: 6.76
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 5.82
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: FAM53B
Fragment: 164-173
Site: 14-3-3_pSer169
Modification: (SEP)169
Sequence: RSSSFSLPSR
Construct: biotin-ttds-RSSSF(pSer)LPSR (HPLC-purified synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Measured dissociation constant (10-6M): 3.4
Standard deviation of dissociation constant (10-6M): 0.5
Experiment method: competitive Fluorescence Polarization
Details of affinity fitting: Competitive binding equation in ProFit
Date of measurement: 2022-01-10
Number of measurements: 3
Measured pKd: 5.46
DOI: 10.1101/2022.12.08.519103

Experiment series 4

Protein A

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)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Measured dissociation constant (10-6M): 5.8
Standard deviation of dissociation constant (10-6M): 1
Experiment method: competitive Fluorescence Polarization
Details of affinity fitting: Competitive binding equation in ProFit
Date of measurement: 2022-01-18
Number of measurements: 3
Measured pKd: 5.23
DOI: 10.1101/2022.12.08.519103

Experiment series 5

Protein A

Protein: FAM53B
Fragment: 263-272
Site: 14-3-3_pSer268
Modification: (SEP)268
Sequence: LSRSRSQPCV
Construct: LSRSR(pSer)QPCV (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: -0.15
Holdup BI standard deviation: 0.51
Immobilized partner concentration (10-6M): 129
P value: 0.3
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: FAM53B
Fragment: 164-173
Site: 14-3-3_pSer169
Modification: (SEP)169
Sequence: RSSSFSLPSR
Construct: RSSSF(pSer)LPSR (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Measured dissociation constant (10-6M): <1
Immobilized partner concentration (10-6M): 115
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 1
Measured pKd: 6
DOI: 10.1101/2022.12.08.519103
Experimental Details

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