Interaction Details


Details of experiments between P31946 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: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta

Protein B

Protein: PI4KB
Fragment: 289-298
Site: 14-3-3_pSer294
Modification: (SEP)294
Sequence: LKRTASNPKV
Construct: LKRTA(pSer)NPKV (crude synthetic peptide)
Average holdup BI: 0.97
Holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 134
P value: 5.12
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 5.36
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: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta

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: -0.12
Holdup BI standard deviation: 0.22
Immobilized partner concentration (10-6M): 134
P value: 0.52
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: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta

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: -0.13
Holdup BI standard deviation: 0.71
Immobilized partner concentration (10-6M): 102
P value: 0.16
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 4

Protein A

Protein: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta

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: 0.78
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 102
P value: 7.68
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.53
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: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta

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: 0.78
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 134
P value: 9.53
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.42
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: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta

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: 0.41
Holdup BI standard deviation: 0.15
Immobilized partner concentration (10-6M): 134
P value: 2.64
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.72
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: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta

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: 0.3
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 102
P value: 5.4
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.63
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: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta

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: 0.41
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 134
P value: 4.79
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.72
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: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta

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: 0.33
Holdup BI standard deviation: 0.09
Immobilized partner concentration (10-6M): 102
P value: 3.73
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.69
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: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta

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: 0.09
Holdup BI standard deviation: 0.09
Immobilized partner concentration (10-6M): 134
P value: 0.98
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: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta

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: -0.05
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 102
P value: 0.88
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))