Interaction Details


Details of experiments between P63104 and O43896
Note that the results of all experiments are listed, regardless of the modification states of the fragments.


Experiment series 1

Protein A

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta

Protein B

Protein: KIF1C
Fragment: 1087-1096
Site: 14-3-3_pSer1092
Modification: (SEP)1092
Sequence: MRRQRSAPDL
Construct: MRRQR(pSer)APDL (crude synthetic peptide)
Average holdup BI: 0.86
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 132
P value: 8.96
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.66
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: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta

Protein B

Protein: KIF1C
Fragment: 669-678
Site: 14-3-3_pSer674
Modification: (SEP)674
Sequence: RLYADSDSGD
Construct: RLYAD(pSer)DSGD (crude synthetic peptide)
Average holdup BI: -0.31
Holdup BI standard deviation: 0.72
Immobilized partner concentration (10-6M): 132
P value: 0.61
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 3

Protein A

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta

Protein B

Protein: KIF1C
Fragment: 1077-1086
Site: 14-3-3_pThr1082
Modification: (TPO)1082
Sequence: RYPPYTTPPR
Construct: RYPPY(pThr)TPPR (crude synthetic peptide)
Average holdup BI: 0.98
Holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 132
P value: 4.94
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 5.56
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: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta

Protein B

Protein: KIF1C
Fragment: 1077-1086
Site: 14-3-3_pThr1082
Modification: (TPO)1082
Sequence: RYPPYTTPPR
Construct: biotin-ttds-RYPPY(pThr)TPPR (HPLC-purified synthetic peptide)
Measured dissociation constant (10-6M): 14
Standard deviation of dissociation constant (10-6M): 1.2
Experiment method: competitive Fluorescence Polarization
Details of affinity fitting: Competitive binding equation in ProFit
Date of measurement: 2022-02-10
Number of measurements: 3
Measured pKd: 4.85
DOI: 10.1101/2022.12.08.519103

Experiment series 5

Protein A

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta

Protein B

Protein: KIF1C
Fragment: 1077-1086
Site: 14-3-3_pThr1082
Modification: (TPO)1082
Sequence: RYPPYTTPPR
Construct: biotin-ttds-RYPPY(pThr)TPPR (HPLC-purified synthetic peptide)
Measured dissociation constant (10-6M): 21
Standard deviation of dissociation constant (10-6M): 2
Experiment method: competitive Fluorescence Polarization
Details of affinity fitting: Competitive binding equation in ProFit
Date of measurement: 2022-01-19
Number of measurements: 3
Measured pKd: 4.68
DOI: 10.1101/2022.12.08.519103

Experiment series 6

Protein A

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta

Protein B

Protein: KIF1C
Fragment: 272-281
Site: 14-3-3_pThr277
Modification: (TPO)277
Sequence: NKSLTTLGKV
Construct: NKSLT(pThr)LGKV (crude synthetic peptide)
Average holdup BI: -0.04
Holdup BI standard deviation: 0.13
Immobilized partner concentration (10-6M): 132
P value: 0.29
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 7

Protein A

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta

Protein B

Protein: KIF1C
Fragment: 484-493
Site: 14-3-3_pThr489
Modification: (TPO)489
Sequence: REDGGTVGVF
Construct: REDGG(pThr)VGVF (crude synthetic peptide)
Average holdup BI: 0.05
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 132
P value: 1.25
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))