Interaction Details


Details of experiments between O94915 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: FRYL
Fragment: 98-107
Site: 14-3-3_pSer103
Modification: (SEP)103
Sequence: RSSTKSKGDE
Construct: RSSTK(pSer)KGDE (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: 0.3
Holdup BI standard deviation: 0.62
Immobilized partner concentration (10-6M): 134
P value: 0.39
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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.08
Holdup BI standard deviation: 0.81
Immobilized partner concentration (10-6M): 129
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 3

Protein A

Protein: FRYL
Fragment: 1530-1539
Site: 14-3-3_pSer1535
Modification: (SEP)1535
Sequence: SRYSSSSGGS
Construct: SRYSS(pSer)SGGS (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: -0.21
Holdup BI standard deviation: 1.81
Immobilized partner concentration (10-6M): 134
P value: 0.08
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 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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Measured dissociation constant (10-6M): <100
Immobilized partner concentration (10-6M): 134
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 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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.02
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 134
P value: 0.56
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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.05
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 129
P value: 0.83
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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.59
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 134
P value: 7.87
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.03
DOI: 10.1101/2022.12.08.519103
Experimental Details

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


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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.56
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 129
P value: 8.51
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.99
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: 1940-1949
Site: 14-3-3_pSer1945
Modification: (SEP)1945
Sequence: NSLRLSLIGD
Construct: NSLRL(pSer)LIGD (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: -0.08
Holdup BI standard deviation: 0.64
Immobilized partner concentration (10-6M): 129
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 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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.13
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 134
P value: 3.44
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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.13
Holdup BI standard deviation: 0.09
Immobilized partner concentration (10-6M): 129
P value: 1.67
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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.94
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 129
P value: 9.76
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 5.1
DOI: 10.1101/2022.12.08.519103
Experimental Details

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


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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.93
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 134
P value: 5.99
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 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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Measured dissociation constant (10-6M): <10
Immobilized partner concentration (10-6M): 129
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 1
Measured pKd: 5
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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Measured dissociation constant (10-6M): <100
Immobilized partner concentration (10-6M): 134
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 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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: -0.4
Holdup BI standard deviation: 2.17
Immobilized partner concentration (10-6M): 134
P value: 0.14
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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.07
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 134
P value: 1.99
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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.04
Holdup BI standard deviation: 0.08
Immobilized partner concentration (10-6M): 129
P value: 0.52
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 19

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: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.02
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 115
P value: 0.43
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 20

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: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.52
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 115
P value: 6.27
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.99
DOI: 10.1101/2022.12.08.519103
Experimental Details

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


Experiment series 21

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: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.17
Holdup BI standard deviation: 0.08
Immobilized partner concentration (10-6M): 115
P value: 2.04
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 22

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: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.12
Holdup BI standard deviation: 0.19
Immobilized partner concentration (10-6M): 115
P value: 0.62
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 23

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: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.92
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 115
P value: 6.96
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 5.03
DOI: 10.1101/2022.12.08.519103
Experimental Details

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


Experiment series 24

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: 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): <100
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: 4
DOI: 10.1101/2022.12.08.519103
Experimental Details

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


Experiment series 25

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: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.13
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 115
P value: 3.11
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))