Interaction Details


Details of experiments between Q9UBF8 and P63104
Note that the results of all experiments are listed, regardless of the modification states of the fragments.


Experiment series 1

Protein A

Protein: PI4KB
Fragment: 420-429
Site: 14-3-3_pSer425
Modification: (SEP)425
Sequence: ENRIRSTRSV
Construct: ENRIR(pSer)TRSV (crude synthetic peptide)

Protein B

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta
Average holdup BI: 0.05
Holdup BI standard deviation: 0.4
Immobilized partner concentration (10-6M): 132
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 2

Protein A

Protein: PI4KB
Fragment: 423-432
Site: 14-3-3_pSer428
Modification: (SEP)428
Sequence: IRSTRSVENL
Construct: IRSTR(pSer)VENL (crude synthetic peptide)

Protein B

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta
Average holdup BI: 0.74
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 132
P value: 7.3
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.34
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: PI4KB
Fragment: 506-515
Site: 14-3-3_pSer511
Modification: (SEP)511
Sequence: IRRRLSEQLA
Construct: IRRRL(pSer)EQLA (crude synthetic peptide)

Protein B

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta
Average holdup BI: 0.28
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 132
P value: 5.3
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.46
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: PI4KB
Fragment: 336-345
Site: 14-3-3_pThr341
Modification: (TPO)341
Sequence: GKRLATLPTK
Construct: GKRLA(pThr)LPTK (crude synthetic peptide)

Protein B

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta
Average holdup BI: 0.29
Holdup BI standard deviation: 0.08
Immobilized partner concentration (10-6M): 132
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: 3.49
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: PI4KB
Fragment: 499-508
Site: 14-3-3_pThr504
Modification: (TPO)504
Sequence: LAHTPTAFKR
Construct: LAHTP(pThr)AFKR (crude synthetic peptide)

Protein B

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta
Average holdup BI: -0.06
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 132
P value: 1.05
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))