Interaction Details


Details of experiments between P31947 and Q9UBF8
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: PI4KB
Fragment: 289-298
Site: 14-3-3_pSer294
Modification: (SEP)294
Sequence: LKRTASNPKV
Construct: LKRTA(pSer)NPKV (crude synthetic peptide)
Measured dissociation constant (10-6M): <10
Immobilized partner concentration (10-6M): 113
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
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 4))

Experiment series 2

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: PI4KB
Fragment: 289-298
Site: 14-3-3_pSer294
Modification: (SEP)294
Sequence: LKRTASNPKV
Construct: LKRTA(pSer)NPKV (crude synthetic peptide)
Measured dissociation constant (10-6M): <10
Immobilized partner concentration (10-6M): 111
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
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 4))

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: PI4KB
Fragment: 420-429
Site: 14-3-3_pSer425
Modification: (SEP)425
Sequence: ENRIRSTRSV
Construct: ENRIR(pSer)TRSV (crude synthetic peptide)
Average holdup BI: -0.01
Holdup BI standard deviation: 0.27
Immobilized partner concentration (10-6M): 113
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 4

Protein A

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

Protein B

Protein: PI4KB
Fragment: 420-429
Site: 14-3-3_pSer425
Modification: (SEP)425
Sequence: ENRIRSTRSV
Construct: ENRIR(pSer)TRSV (crude synthetic peptide)
Average holdup BI: -0.15
Holdup BI standard deviation: 0.49
Immobilized partner concentration (10-6M): 142
P value: 0.35
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: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: PI4KB
Fragment: 420-429
Site: 14-3-3_pSer425
Modification: (SEP)425
Sequence: ENRIRSTRSV
Construct: ENRIR(pSer)TRSV (crude synthetic peptide)
Average holdup BI: -0.14
Holdup BI standard deviation: 0.82
Immobilized partner concentration (10-6M): 111
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 6

Protein A

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

Protein B

Protein: PI4KB
Fragment: 423-432
Site: 14-3-3_pSer428
Modification: (SEP)428
Sequence: IRSTRSVENL
Construct: IRSTR(pSer)VENL (crude synthetic peptide)
Average holdup BI: 0.45
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 142
P value: 5.91
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.76
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: PI4KB
Fragment: 423-432
Site: 14-3-3_pSer428
Modification: (SEP)428
Sequence: IRSTRSVENL
Construct: IRSTR(pSer)VENL (crude synthetic peptide)
Average holdup BI: 0.39
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 113
P value: 7.33
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.74
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
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: PI4KB
Fragment: 423-432
Site: 14-3-3_pSer428
Modification: (SEP)428
Sequence: IRSTRSVENL
Construct: IRSTR(pSer)VENL (crude synthetic peptide)
Average holdup BI: 0.46
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 111
P value: 6.36
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.88
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: PI4KB
Fragment: 506-515
Site: 14-3-3_pSer511
Modification: (SEP)511
Sequence: IRRRLSEQLA
Construct: IRRRL(pSer)EQLA (crude synthetic peptide)
Average holdup BI: -0.08
Holdup BI standard deviation: 0.09
Immobilized partner concentration (10-6M): 113
P value: 0.93
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: PI4KB
Fragment: 506-515
Site: 14-3-3_pSer511
Modification: (SEP)511
Sequence: IRRRLSEQLA
Construct: IRRRL(pSer)EQLA (crude synthetic peptide)
Average holdup BI: 0.08
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 142
P value: 1.54
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 11

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: PI4KB
Fragment: 506-515
Site: 14-3-3_pSer511
Modification: (SEP)511
Sequence: IRRRLSEQLA
Construct: IRRRL(pSer)EQLA (crude synthetic peptide)
Average holdup BI: 0.08
Holdup BI standard deviation: 0.08
Immobilized partner concentration (10-6M): 111
P value: 1.1
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 12

Protein A

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

Protein B

Protein: PI4KB
Fragment: 336-345
Site: 14-3-3_pThr341
Modification: (TPO)341
Sequence: GKRLATLPTK
Construct: GKRLA(pThr)LPTK (crude synthetic peptide)
Average holdup BI: 0.31
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 142
P value: 4.03
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.49
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: PI4KB
Fragment: 336-345
Site: 14-3-3_pThr341
Modification: (TPO)341
Sequence: GKRLATLPTK
Construct: GKRLA(pThr)LPTK (crude synthetic peptide)
Average holdup BI: 0.21
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 113
P value: 3.02
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.36
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
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: PI4KB
Fragment: 336-345
Site: 14-3-3_pThr341
Modification: (TPO)341
Sequence: GKRLATLPTK
Construct: GKRLA(pThr)LPTK (crude synthetic peptide)
Average holdup BI: 0.15
Holdup BI standard deviation: 0.13
Immobilized partner concentration (10-6M): 111
P value: 1.11
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 15

Protein A

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

Protein B

Protein: PI4KB
Fragment: 499-508
Site: 14-3-3_pThr504
Modification: (TPO)504
Sequence: LAHTPTAFKR
Construct: LAHTP(pThr)AFKR (crude synthetic peptide)
Average holdup BI: -0.03
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 113
P value: 1.43
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 16

Protein A

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

Protein B

Protein: PI4KB
Fragment: 499-508
Site: 14-3-3_pThr504
Modification: (TPO)504
Sequence: LAHTPTAFKR
Construct: LAHTP(pThr)AFKR (crude synthetic peptide)
Average holdup BI: -0.1
Holdup BI standard deviation: 0.09
Immobilized partner concentration (10-6M): 142
P value: 1.23
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 17

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: PI4KB
Fragment: 499-508
Site: 14-3-3_pThr504
Modification: (TPO)504
Sequence: LAHTPTAFKR
Construct: LAHTP(pThr)AFKR (crude synthetic peptide)
Average holdup BI: -0.03
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 111
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))