Interaction Details


Details of experiments between O94915 and Q04917
Note that the results of all experiments are listed, regardless of the modification states of the fragments.



Experiment series 1

Protein A

Protein: FRYL
Fragment: 98-107
Site: 14-3-3_pSer103
Modification: (SEP)103
Sequence: RSSTKSKGDE
Construct: RSSTK(pSer)KGDE (crude synthetic peptide)

Protein B

Protein: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta
Average holdup BI: -0.29
Holdup BI standard deviation: 0.67
Immobilized partner concentration (10-6M): 130
P value: 0.41
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 2

Protein A

Protein: FRYL
Fragment: 1909-1918
Site: 14-3-3_pSer1914
Modification: (SEP)1914
Sequence: AANRKSTGQL
Construct: AANRK(pSer)TGQL (crude synthetic peptide)

Protein B

Protein: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta
Average holdup BI: -0.07
Holdup BI standard deviation: 0.47
Immobilized partner concentration (10-6M): 130
P value: 0.09
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: FRYL
Fragment: 1930-1939
Site: 14-3-3_pSer1935
Modification: (SEP)1935
Sequence: YLGYNSNARS
Construct: YLGYN(pSer)NARS (crude synthetic peptide)

Protein B

Protein: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta
Average holdup BI: 0.07
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 130
P value: 1.21
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 4

Protein A

Protein: FRYL
Fragment: 1936-1945
Site: 14-3-3_pSer1941
Modification: (SEP)1941
Sequence: NARSNSLRLS
Construct: NARSN(pSer)LRLS (crude synthetic peptide)

Protein B

Protein: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta
Average holdup BI: 0.5
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 130
P value: 6.41
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.88
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: FRYL
Fragment: 1940-1949
Site: 14-3-3_pSer1945
Modification: (SEP)1945
Sequence: NSLRLSLIGD
Construct: NSLRL(pSer)LIGD (crude synthetic peptide)

Protein B

Protein: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta
Average holdup BI: -0.04
Holdup BI standard deviation: 0.62
Immobilized partner concentration (10-6M): 130
P value: 0.05
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: FRYL
Fragment: 1967-1976
Site: 14-3-3_pSer1972
Modification: (SEP)1972
Sequence: INHSSSLART
Construct: INHSS(pSer)LART (crude synthetic peptide)

Protein B

Protein: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta
Average holdup BI: 0.16
Holdup BI standard deviation: 0.08
Immobilized partner concentration (10-6M): 130
P value: 2.15
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 7

Protein A

Protein: FRYL
Fragment: 1973-1982
Site: 14-3-3_pSer1978
Modification: (SEP)1978
Sequence: LARTRSLSSL
Construct: LARTR(pSer)LSSL (crude synthetic peptide)

Protein B

Protein: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta
Average holdup BI: 0.91
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 130
P value: 8.73
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.91
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: FRYL
Fragment: 1954-1963
Site: 14-3-3_pThr1959
Modification: (TPO)1959
Sequence: RRRSNTLDIM
Construct: RRRSN(pThr)LDIM (crude synthetic peptide)

Protein B

Protein: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta
Average holdup BI: 0.84
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 130
P value: 2.55
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.61
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))


Experiment series 9

Protein A

Protein: FRYL
Fragment: 477-486
Site: 14-3-3_pThr482
Modification: (TPO)482
Sequence: LPSGNTLRVK
Construct: LPSGN(pThr)LRVK (crude synthetic peptide)

Protein B

Protein: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta
Average holdup BI: 0.03
Holdup BI standard deviation: 0.1
Immobilized partner concentration (10-6M): 130
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))