Interaction Details


Details of experiments between O94915 and P62258
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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: -0.25
Holdup BI standard deviation: 0.5
Immobilized partner concentration (10-6M): 116
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 2

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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: -0.16
Holdup BI standard deviation: 0.51
Immobilized partner concentration (10-6M): 143
P value: 0.27
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: 1909-1918
Site: 14-3-3_pSer1914
Modification: (SEP)1914
Sequence: AANRKSTGQL
Construct: AANRK(pSer)TGQL (crude synthetic peptide)

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Measured dissociation constant (10-6M): <100
Immobilized partner concentration (10-6M): 116
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 1
Measured pKd: 4
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: FRYL
Fragment: 1909-1918
Site: 14-3-3_pSer1914
Modification: (SEP)1914
Sequence: AANRKSTGQL
Construct: AANRK(pSer)TGQL (crude synthetic peptide)

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: -0.15
Holdup BI standard deviation: 0.85
Immobilized partner concentration (10-6M): 143
P value: 0.11
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 5

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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: -0.02
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 116
P value: 0.53
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 6

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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.05
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 143
P value: 0.86
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: 1936-1945
Site: 14-3-3_pSer1941
Modification: (SEP)1941
Sequence: NARSNSLRLS
Construct: NARSN(pSer)LRLS (crude synthetic peptide)

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.22
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 143
P value: 3.82
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.3
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: 1936-1945
Site: 14-3-3_pSer1941
Modification: (SEP)1941
Sequence: NARSNSLRLS
Construct: NARSN(pSer)LRLS (crude synthetic peptide)

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.24
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 116
P value: 4.77
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: FRYL
Fragment: 1940-1949
Site: 14-3-3_pSer1945
Modification: (SEP)1945
Sequence: NSLRLSLIGD
Construct: NSLRL(pSer)LIGD (crude synthetic peptide)

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: -0.22
Holdup BI standard deviation: 0.25
Immobilized partner concentration (10-6M): 116
P value: 0.89
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 10

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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.09
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 116
P value: 1.78
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: FRYL
Fragment: 1967-1976
Site: 14-3-3_pSer1972
Modification: (SEP)1972
Sequence: INHSSSLART
Construct: INHSS(pSer)LART (crude synthetic peptide)

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.01
Holdup BI standard deviation: 0.14
Immobilized partner concentration (10-6M): 143
P value: 0.07
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 12

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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.79
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 116
P value: 5.52
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.52
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: FRYL
Fragment: 1973-1982
Site: 14-3-3_pSer1978
Modification: (SEP)1978
Sequence: LARTRSLSSL
Construct: LARTR(pSer)LSSL (crude synthetic peptide)

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.82
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 143
P value: 9.12
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.51
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 14

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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.76
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 143
P value: 4.63
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.34
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 15

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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: -0.2
Holdup BI standard deviation: 1.29
Immobilized partner concentration (10-6M): 116
P value: 0.13
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 16

Protein A

Protein: FRYL
Fragment: 2801-2810
Site: 14-3-3_pThr2806
Modification: (TPO)2806
Sequence: DDCKRTFGAK
Construct: DDCKR(pThr)FGAK (crude synthetic peptide)

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.12
Holdup BI standard deviation: 1.59
Immobilized partner concentration (10-6M): 116
P value: 0.05
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 17

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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.02
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 116
P value: 0.68
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 18

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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: -0.01
Holdup BI standard deviation: 0.1
Immobilized partner concentration (10-6M): 143
P value: 0.08
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))