Interaction Details


Details of experiments between P31946 and O60333
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: KIF1B
Fragment: 1048-1057
Site: 14-3-3_pSer1053
Modification: (SEP)1053
Sequence: RWRSNSLNNG
Construct: RWRSN(pSer)LNNG (crude synthetic peptide)
Average holdup BI: 0.8
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 102
P value: 7.25
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.6
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: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta

Protein B

Protein: KIF1B
Fragment: 1048-1057
Site: 14-3-3_pSer1053
Modification: (SEP)1053
Sequence: RWRSNSLNNG
Construct: RWRSN(pSer)LNNG (crude synthetic peptide)
Average holdup BI: 0.84
Holdup BI standard deviation: 0.11
Immobilized partner concentration (10-6M): 134
P value: 5.11
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.58
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: KIF1B
Fragment: 1052-1061
Site: 14-3-3_pSer1057
Modification: (SEP)1057
Sequence: TRSGLSLEEL
Construct: TRSGL(pSer)LEEL (crude synthetic peptide)
Average holdup BI: 0.13
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 134
P value: 2.18
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: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta

Protein B

Protein: KIF1B
Fragment: 1052-1061
Site: 14-3-3_pSer1057
Modification: (SEP)1057
Sequence: TRSGLSLEEL
Construct: TRSGL(pSer)LEEL (crude synthetic peptide)
Average holdup BI: 0.18
Holdup BI standard deviation: 0.13
Immobilized partner concentration (10-6M): 102
P value: 1.56
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: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta

Protein B

Protein: KIF1B
Fragment: 1136-1145
Site: 14-3-3_pSer1141
Modification: (SEP)1141
Sequence: MRRQFSAPNL
Construct: MRRQF(pSer)APNL (crude synthetic peptide)
Average holdup BI: 0.98
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 134
P value: 2.8
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 5.65
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: KIF1B
Fragment: 1136-1145
Site: 14-3-3_pSer1141
Modification: (SEP)1141
Sequence: MRRQFSAPNL
Construct: MRRQF(pSer)APNL (crude synthetic peptide)
Average holdup BI: 0.94
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 102
P value: 6.18
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 5.18
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: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta

Protein B

Protein: KIF1B
Fragment: 1482-1491
Site: 14-3-3_pSer1487
Modification: (SEP)1487
Sequence: RPRGDSLILE
Construct: RPRGD(pSer)LILE (crude synthetic peptide)
Average holdup BI: 0.06
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 134
P value: 1.47
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: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta

Protein B

Protein: KIF1B
Fragment: 1482-1491
Site: 14-3-3_pSer1487
Modification: (SEP)1487
Sequence: RPRGDSLILE
Construct: RPRGD(pSer)LILE (crude synthetic peptide)
Average holdup BI: 0.03
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 102
P value: 0.46
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: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta

Protein B

Protein: KIF1B
Fragment: 658-667
Site: 14-3-3_pSer663
Modification: (SEP)663
Sequence: RLDADSDSGD
Construct: RLDAD(pSer)DSGD (crude synthetic peptide)
Average holdup BI: -0.18
Holdup BI standard deviation: 0.17
Immobilized partner concentration (10-6M): 134
P value: 1.33
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: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta

Protein B

Protein: KIF1B
Fragment: 887-896
Site: 14-3-3_pSer892
Modification: (SEP)892
Sequence: FKLVGSSPIF
Construct: FKLVG(pSer)SPIF (crude synthetic peptide)
Average holdup BI: 0.85
Holdup BI standard deviation: 0.08
Immobilized partner concentration (10-6M): 102
P value: 2.62
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.74
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: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta

Protein B

Protein: KIF1B
Fragment: 887-896
Site: 14-3-3_pSer892
Modification: (SEP)892
Sequence: FKLVGSSPIF
Construct: FKLVG(pSer)SPIF (crude synthetic peptide)
Average holdup BI: 0.87
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 134
P value: 2.76
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.7
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: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta

Protein B

Protein: KIF1B
Fragment: 517-526
Site: 14-3-3_pThr522
Modification: (TPO)522
Sequence: REDGGTLGVF
Construct: REDGG(pThr)LGVF (crude synthetic peptide)
Average holdup BI: 0.07
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 134
P value: 1.65
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 13

Protein A

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

Protein B

Protein: KIF1B
Fragment: 517-526
Site: 14-3-3_pThr522
Modification: (TPO)522
Sequence: REDGGTLGVF
Construct: REDGG(pThr)LGVF (crude synthetic peptide)
Average holdup BI: 0.12
Holdup BI standard deviation: 0.16
Immobilized partner concentration (10-6M): 102
P value: 0.76
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))