Interaction Details


Details of experiments between P62258 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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

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: 0.76
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 143
P value: 7.4
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 2

Protein A

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

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: 0.71
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 116
P value: 4.86
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.32
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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

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: -0.29
Holdup BI standard deviation: 0.7
Immobilized partner concentration (10-6M): 143
P value: 0.58
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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

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: 0.02
Immobilized partner concentration (10-6M): 116
P value: 6.52
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 5.55
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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

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): 17
Standard deviation of dissociation constant (10-6M): 3
Experiment method: competitive Fluorescence Polarization
Details of affinity fitting: Competitive binding equation in ProFit
Date of measurement: 2022-02-09
Number of measurements: 3
Measured pKd: 4.76
DOI: 10.1101/2022.12.08.519103

Experiment series 6

Protein A

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

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: 0.89
Holdup BI standard deviation: 0.1
Immobilized partner concentration (10-6M): 143
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: 4.76
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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

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: -0.02
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 116
P value: 0.32
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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

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: -0.03
Holdup BI standard deviation: 0.12
Immobilized partner concentration (10-6M): 143
P value: 0.24
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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

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): 116
P value: 1.51
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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

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: 0.01
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 143
P value: 0.26
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))