Interaction Details


Details of experiments between P31947 and Q9H4L5
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: OSBPL3
Fragment: 246-255
Site: 14-3-3_pSer251
Modification: (SEP)251
Sequence: LHRTYSAPAI
Construct: LHRTY(pSer)APAI (crude synthetic peptide)
Average holdup BI: 0.81
Holdup BI standard deviation: 0.1
Immobilized partner concentration (10-6M): 113
P value: 2.6
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.58
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: OSBPL3
Fragment: 246-255
Site: 14-3-3_pSer251
Modification: (SEP)251
Sequence: LHRTYSAPAI
Construct: LHRTY(pSer)APAI (crude synthetic peptide)
Average holdup BI: 0.67
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 142
P value: 6.26
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.15
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: OSBPL3
Fragment: 299-308
Site: 14-3-3_pSer304
Modification: (SEP)304
Sequence: VRLHSSNPNL
Construct: VRLHS(pSer)NPNL (crude synthetic peptide)
Average holdup BI: 0.27
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 142
P value: 3.69
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.41
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: OSBPL3
Fragment: 299-308
Site: 14-3-3_pSer304
Modification: (SEP)304
Sequence: VRLHSSNPNL
Construct: VRLHS(pSer)NPNL (crude synthetic peptide)
Average holdup BI: 0.21
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 113
P value: 4.16
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.37
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: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: OSBPL3
Fragment: 299-308
Site: 14-3-3_pSer304
Modification: (SEP)304
Sequence: VRLHSSNPNL
Construct: VRLHS(pSer)NPNL (crude synthetic peptide)
Average holdup BI: 0.16
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 111
P value: 5.08
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 6

Protein A

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

Protein B

Protein: OSBPL3
Fragment: 315-324
Site: 14-3-3_pSer320
Modification: (SEP)320
Sequence: EEKNYSDGSE
Construct: EEKNY(pSer)DGSE (crude synthetic peptide)
Average holdup BI: 0.93
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 113
P value: 1.41
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 5.06
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: OSBPL3
Fragment: 29-38
Site: 14-3-3_pSer34
Modification: (SEP)34
Sequence: GSRQDSWEVV
Construct: GSRQD(pSer)WEVV (crude synthetic peptide)
Average holdup BI: 0.06
Holdup BI standard deviation: 0.13
Immobilized partner concentration (10-6M): 113
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 8

Protein A

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

Protein B

Protein: OSBPL3
Fragment: 29-38
Site: 14-3-3_pSer34
Modification: (SEP)34
Sequence: GSRQDSWEVV
Construct: GSRQD(pSer)WEVV (crude synthetic peptide)
Average holdup BI: 0.03
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 142
P value: 0.5
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: OSBPL3
Fragment: 29-38
Site: 14-3-3_pSer34
Modification: (SEP)34
Sequence: GSRQDSWEVV
Construct: GSRQD(pSer)WEVV (crude synthetic peptide)
Average holdup BI: -0.01
Holdup BI standard deviation: 0.26
Immobilized partner concentration (10-6M): 111
P value: 0.03
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: OSBPL3
Fragment: 432-441
Site: 14-3-3_pSer437
Modification: (SEP)437
Sequence: LVHQLSNESR
Construct: LVHQL(pSer)NESR (crude synthetic peptide)
Average holdup BI: -0.01
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 113
P value: 0.11
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))

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: OSBPL3
Fragment: 432-441
Site: 14-3-3_pSer437
Modification: (SEP)437
Sequence: LVHQLSNESR
Construct: LVHQL(pSer)NESR (crude synthetic peptide)
Average holdup BI: 0
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 142
P value: 0.1
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: OSBPL3
Fragment: 432-441
Site: 14-3-3_pSer437
Modification: (SEP)437
Sequence: LVHQLSNESR
Construct: LVHQL(pSer)NESR (crude synthetic peptide)
Average holdup BI: -0.06
Holdup BI standard deviation: 0.11
Immobilized partner concentration (10-6M): 111
P value: 0.54
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))