Interaction Details


Details of experiments between P31947 and Q15032
Note that the results of all experiments are listed, regardless of the modification states of the fragments.


Experiment series 1

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: R3HDM1
Fragment: 1092-1101
Site: 14-3-3_pSer1097
Modification: (SEP)1097
Sequence: ILERASSQ--
Construct: FHILERA(pSer)SQ (crude synthetic peptide)
Average holdup BI: 0.24
Holdup BI standard deviation: 0.13
Immobilized partner concentration (10-6M): 142
P value: 1.93
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.35
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: R3HDM1
Fragment: 1092-1101
Site: 14-3-3_pSer1097
Modification: (SEP)1097
Sequence: ILERASSQ--
Construct: FHILERA(pSer)SQ (crude synthetic peptide)
Average holdup BI: 0.08
Holdup BI standard deviation: 0.13
Immobilized partner concentration (10-6M): 113
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 3

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: R3HDM1
Fragment: 1092-1101
Site: 14-3-3_pSer1097
Modification: (SEP)1097
Sequence: ILERASSQ--
Construct: FHILERA(pSer)SQ (crude synthetic peptide)
Average holdup BI: 0.39
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 111
P value: 5.83
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.76
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: R3HDM1
Fragment: 1093-1102
Site: 14-3-3_pSer1098
Modification: (SEP)1098
Sequence: LERASSQ---
Construct: FHILERAS(pSer)Q (crude synthetic peptide)
Average holdup BI: 0.39
Holdup BI standard deviation: 0.14
Immobilized partner concentration (10-6M): 142
P value: 2.96
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.65
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: R3HDM1
Fragment: 1093-1102
Site: 14-3-3_pSer1098
Modification: (SEP)1098
Sequence: LERASSQ---
Construct: FHILERAS(pSer)Q (crude synthetic peptide)
Average holdup BI: 0.32
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 113
P value: 4.8
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.62
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: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: R3HDM1
Fragment: 1093-1102
Site: 14-3-3_pSer1098
Modification: (SEP)1098
Sequence: LERASSQ---
Construct: FHILERAS(pSer)Q (crude synthetic peptide)
Average holdup BI: 0.38
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 111
P value: 5.53
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.73
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 7

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: R3HDM1
Fragment: 137-146
Site: 14-3-3_pSer142
Modification: (SEP)142
Sequence: LSRDSSQEYT
Construct: LSRDS(pSer)QEYT (crude synthetic peptide)
Average holdup BI: 0.01
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 113
P value: 0.25
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 8

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: R3HDM1
Fragment: 137-146
Site: 14-3-3_pSer142
Modification: (SEP)142
Sequence: LSRDSSQEYT
Construct: LSRDS(pSer)QEYT (crude synthetic peptide)
Average holdup BI: 0
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 142
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))

Experiment series 9

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: R3HDM1
Fragment: 137-146
Site: 14-3-3_pSer142
Modification: (SEP)142
Sequence: LSRDSSQEYT
Construct: LSRDS(pSer)QEYT (crude synthetic peptide)
Average holdup BI: -0.02
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 111
P value: 0.47
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: R3HDM1
Fragment: 275-284
Site: 14-3-3_pSer280
Modification: (SEP)280
Sequence: KDDRRSKSIE
Construct: KDDRR(pSer)KSIE (crude synthetic peptide)
Average holdup BI: -0.08
Holdup BI standard deviation: 0.16
Immobilized partner concentration (10-6M): 113
P value: 0.49
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: R3HDM1
Fragment: 275-284
Site: 14-3-3_pSer280
Modification: (SEP)280
Sequence: KDDRRSKSIE
Construct: KDDRR(pSer)KSIE (crude synthetic peptide)
Average holdup BI: 0.02
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 142
P value: 0.26
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: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: R3HDM1
Fragment: 275-284
Site: 14-3-3_pSer280
Modification: (SEP)280
Sequence: KDDRRSKSIE
Construct: KDDRR(pSer)KSIE (crude synthetic peptide)
Average holdup BI: 0.08
Holdup BI standard deviation: 0.15
Immobilized partner concentration (10-6M): 111
P value: 0.48
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 13

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: R3HDM1
Fragment: 294-303
Site: 14-3-3_pSer299
Modification: (SEP)299
Sequence: RDRIFSQDSL
Construct: RDRIF(pSer)QDSL (crude synthetic peptide)
Average holdup BI: -0.44
Holdup BI standard deviation: 0.3
Immobilized partner concentration (10-6M): 113
P value: 2.07
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 14

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: R3HDM1
Fragment: 294-303
Site: 14-3-3_pSer299
Modification: (SEP)299
Sequence: RDRIFSQDSL
Construct: RDRIF(pSer)QDSL (crude synthetic peptide)
Average holdup BI: 0.01
Holdup BI standard deviation: 0.1
Immobilized partner concentration (10-6M): 142
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 15

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: R3HDM1
Fragment: 294-303
Site: 14-3-3_pSer299
Modification: (SEP)299
Sequence: RDRIFSQDSL
Construct: RDRIF(pSer)QDSL (crude synthetic peptide)
Average holdup BI: -0.02
Holdup BI standard deviation: 0.12
Immobilized partner concentration (10-6M): 111
P value: 0.1
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: R3HDM1
Fragment: 350-359
Site: 14-3-3_pSer355
Modification: (SEP)355
Sequence: NELKYSEPRP
Construct: NELKY(pSer)EPRP (crude synthetic peptide)
Average holdup BI: 0.08
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 113
P value: 1.17
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: R3HDM1
Fragment: 350-359
Site: 14-3-3_pSer355
Modification: (SEP)355
Sequence: NELKYSEPRP
Construct: NELKY(pSer)EPRP (crude synthetic peptide)
Average holdup BI: 0.04
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 142
P value: 0.54
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 18

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: R3HDM1
Fragment: 350-359
Site: 14-3-3_pSer355
Modification: (SEP)355
Sequence: NELKYSEPRP
Construct: NELKY(pSer)EPRP (crude synthetic peptide)
Average holdup BI: 0.01
Holdup BI standard deviation: 0.08
Immobilized partner concentration (10-6M): 111
P value: 0.1
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 19

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: R3HDM1
Fragment: 376-385
Site: 14-3-3_pSer381
Modification: (SEP)381
Sequence: VTKASSFSGI
Construct: VTKAS(pSer)FSGI (crude synthetic peptide)
Average holdup BI: 0.64
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 113
P value: 5.84
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.19
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: R3HDM1
Fragment: 376-385
Site: 14-3-3_pSer381
Modification: (SEP)381
Sequence: VTKASSFSGI
Construct: VTKAS(pSer)FSGI (crude synthetic peptide)
Average holdup BI: 0.61
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 142
P value: 6.35
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.04
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 21

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: R3HDM1
Fragment: 376-385
Site: 14-3-3_pSer381
Modification: (SEP)381
Sequence: VTKASSFSGI
Construct: VTKAS(pSer)FSGI (crude synthetic peptide)
Average holdup BI: 0.61
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 111
P value: 5.68
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.15
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: R3HDM1
Fragment: 388-397
Site: 14-3-3_pSer393
Modification: (SEP)393
Sequence: LTRGDSSGSS
Construct: LTRGD(pSer)SGSS (crude synthetic peptide)
Average holdup BI: 0.11
Holdup BI standard deviation: 1.29
Immobilized partner concentration (10-6M): 113
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 23

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: R3HDM1
Fragment: 388-397
Site: 14-3-3_pSer393
Modification: (SEP)393
Sequence: LTRGDSSGSS
Construct: LTRGD(pSer)SGSS (crude synthetic peptide)
Average holdup BI: -0.01
Holdup BI standard deviation: 0.87
Immobilized partner concentration (10-6M): 111
P value: 0.01
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))