Interaction Details

Details of experiments between P33176 and P61981
Note that the results of all experiments are listed, regardless of the modification states of the fragments.

Experiment series 1

Protein A

Protein: KIF5B
Fragment: 149-158
Site: 14-3-3_pSer154
Modification: (SEP)154
Sequence: SKTNLSVHED
Construct: SKTNL(pSer)VHED (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

Average holdup BI: -0.1

Holdup BI standard deviation: 0.08

Immobilized partner concentration (10^-6M): 134

P value: 1.4

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 2

Protein A

Protein: KIF5B
Fragment: 149-158
Site: 14-3-3_pSer154
Modification: (SEP)154
Sequence: SKTNLSVHED
Construct: SKTNL(pSer)VHED (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

Average holdup BI: -0.03

Holdup BI standard deviation: 0.04

Immobilized partner concentration (10^-6M): 129

P value: 0.66

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: KIF5B
Fragment: 933-942
Site: 14-3-3_pSer938
Modification: (SEP)938
Sequence: SPTHPSAIRG
Construct: SPTHP(pSer)AIRG (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

Average holdup BI: 0.06

Holdup BI standard deviation: 0.14

Immobilized partner concentration (10^-6M): 134

P value: 0.39

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: KIF5B
Fragment: 933-942
Site: 14-3-3_pSer938
Modification: (SEP)938
Sequence: SPTHPSAIRG
Construct: SPTHP(pSer)AIRG (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

Average holdup BI: -0.02

Holdup BI standard deviation: 0.06

Immobilized partner concentration (10^-6M): 129

P value: 0.36

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: KIF5B
Fragment: 945-954
Site: 14-3-3_pSer950
Modification: (SEP)950
Sequence: AFVQNSQPVA
Construct: AFVQN(pSer)QPVA (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

Average holdup BI: 0.21

Holdup BI standard deviation: 0.03

Immobilized partner concentration (10^-6M): 134

P value: 5.22

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 6

Protein A

Protein: KIF5B
Fragment: 945-954
Site: 14-3-3_pSer950
Modification: (SEP)950
Sequence: AFVQNSQPVA
Construct: AFVQN(pSer)QPVA (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

Average holdup BI: 0.2

Holdup BI standard deviation: 0.03

Immobilized partner concentration (10^-6M): 129

P value: 5.08

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: KIF5B
Fragment: 392-401
Site: 14-3-3_pThr397
Modification: (TPO)397
Sequence: VDKDITLTND
Construct: VDKDI(pThr)LTND (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

Average holdup BI: 0.06

Holdup BI standard deviation: 0.03

Immobilized partner concentration (10^-6M): 134

P value: 1.86

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: KIF5B
Fragment: 392-401
Site: 14-3-3_pThr397
Modification: (TPO)397
Sequence: VDKDITLTND
Construct: VDKDI(pThr)LTND (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

Average holdup BI: -0.02

Holdup BI standard deviation: 0.07

Immobilized partner concentration (10^-6M): 129

P value: 0.22

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: KIF5B
Fragment: 446-455
Site: 14-3-3_pThr451
Modification: (TPO)451
Sequence: VEKLKTQMLD
Construct: VEKLK(pThr)QMLD (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

Average holdup BI: -0.14

Holdup BI standard deviation: 0.24

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 10

Protein A

Protein: KIF5B
Fragment: 446-455
Site: 14-3-3_pThr451
Modification: (TPO)451
Sequence: VEKLKTQMLD
Construct: VEKLK(pThr)QMLD (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

Average holdup BI: 0.18

Holdup BI standard deviation: 0.42

Immobilized partner concentration (10^-6M): 129

P value: 0.36

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 11

Protein A

Protein: KIF5B
Fragment: 149-158
Site: 14-3-3_pSer154
Modification: (SEP)154
Sequence: SKTNLSVHED
Construct: SKTNL(pSer)VHED (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

Average holdup BI: -0.04

Holdup BI standard deviation: 0.12

Immobilized partner concentration (10^-6M): 115

P value: 0.27

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 12

Protein A

Protein: KIF5B
Fragment: 933-942
Site: 14-3-3_pSer938
Modification: (SEP)938
Sequence: SPTHPSAIRG
Construct: SPTHP(pSer)AIRG (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

Average holdup BI: 0.05

Holdup BI standard deviation: 0.14

Immobilized partner concentration (10^-6M): 115

P value: 0.28

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: KIF5B
Fragment: 945-954
Site: 14-3-3_pSer950
Modification: (SEP)950
Sequence: AFVQNSQPVA
Construct: AFVQN(pSer)QPVA (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

Average holdup BI: 0.09

Holdup BI standard deviation: 0.05

Immobilized partner concentration (10^-6M): 115

P value: 1.81

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 14

Protein A

Protein: KIF5B
Fragment: 392-401
Site: 14-3-3_pThr397
Modification: (TPO)397
Sequence: VDKDITLTND
Construct: VDKDI(pThr)LTND (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

Average holdup BI: -0.1

Holdup BI standard deviation: 0.08

Immobilized partner concentration (10^-6M): 115

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 15

Protein A

Protein: KIF5B
Fragment: 446-455
Site: 14-3-3_pThr451
Modification: (TPO)451
Sequence: VEKLKTQMLD
Construct: VEKLK(pThr)QMLD (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

Average holdup BI: 0.13

Holdup BI standard deviation: 0.55

Immobilized partner concentration (10^-6M): 115

P value: 0.13

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))