Interaction Details

Details of experiments between Q02241 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: KIF23
Fragment: 155-164
Site: 14-3-3_pSer160
Modification: (SEP)160
Sequence: SNDRNSMDIQ
Construct: SNDRN(pSer)MDIQ (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: 0.22

Holdup BI standard deviation: 0.22

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

P value: 0.94

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: KIF23
Fragment: 155-164
Site: 14-3-3_pSer160
Modification: (SEP)160
Sequence: SNDRNSMDIQ
Construct: SNDRN(pSer)MDIQ (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: -0.08

Holdup BI standard deviation: 0.09

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

P value: 1.09

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: KIF23
Fragment: 13-22
Site: 14-3-3_pSer18
Modification: (SEP)18
Sequence: TVKKGSQTNL
Construct: TVKKG(pSer)QTNL (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: -0.17

Holdup BI standard deviation: 1.18

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

P value: 0.09

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: KIF23
Fragment: 862-871
Site: 14-3-3_pSer867
Modification: (SEP)867
Sequence: HQELASDGEI
Construct: HQELA(pSer)DGEI (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: 0.23

Holdup BI standard deviation: 0.88

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

P value: 0.14

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 5

Protein A

Protein: KIF23
Fragment: 919-928
Site: 14-3-3_pSer924
Modification: (SEP)924
Sequence: PDGAESEWTD
Construct: PDGAE(pSer)EWTD (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: 1.1

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

P value: 0.01

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: KIF23
Fragment: 938-947
Site: 14-3-3_pSer943
Modification: (SEP)943
Sequence: EMRAGSQLGP
Construct: EMRAG(pSer)QLGP (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: 0.23

Holdup BI standard deviation: 0.16

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 7

Protein A

Protein: KIF23
Fragment: 938-947
Site: 14-3-3_pSer943
Modification: (SEP)943
Sequence: EMRAGSQLGP
Construct: EMRAG(pSer)QLGP (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: 0.07

Holdup BI standard deviation: 0.06

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

P value: 1.4

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 8

Protein A

Protein: KIF23
Fragment: 892-901
Site: 14-3-3_pThr897
Modification: (TPO)897
Sequence: FTDIETLKQE
Construct: FTDIE(pThr)LKQE (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: -0.04

Holdup BI standard deviation: 0.06

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

P value: 0.61

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 9

Protein A

Protein: KIF23
Fragment: 892-901
Site: 14-3-3_pThr897
Modification: (TPO)897
Sequence: FTDIETLKQE
Construct: FTDIE(pThr)LKQE (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: 0.02

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

P value: 1.83

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 10

Protein A

Protein: KIF23
Fragment: 922-931
Site: 14-3-3_pThr927
Modification: (TPO)927
Sequence: AESEWTDVET
Construct: AESEW(pThr)DVET (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: 0.04

Holdup BI standard deviation: 0.64

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

P value: 0.04

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: KIF23
Fragment: 155-164
Site: 14-3-3_pSer160
Modification: (SEP)160
Sequence: SNDRNSMDIQ
Construct: SNDRN(pSer)MDIQ (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: -0.08

Holdup BI standard deviation: 0.72

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

P value: 0.08

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: KIF23
Fragment: 677-686
Site: 14-3-3_pSer682
Modification: (SEP)682
Sequence: KVTQRSVSPS
Construct: KVTQR(pSer)VSPS (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: 0.63

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

P value: 0.1

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: KIF23
Fragment: 862-871
Site: 14-3-3_pSer867
Modification: (SEP)867
Sequence: HQELASDGEI
Construct: HQELA(pSer)DGEI (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: -0.14

Holdup BI standard deviation: 2.43

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

P value: 0.04

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: KIF23
Fragment: 938-947
Site: 14-3-3_pSer943
Modification: (SEP)943
Sequence: EMRAGSQLGP
Construct: EMRAG(pSer)QLGP (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: 0.45

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

P value: 0.08

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: KIF23
Fragment: 892-901
Site: 14-3-3_pThr897
Modification: (TPO)897
Sequence: FTDIETLKQE
Construct: FTDIE(pThr)LKQE (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: -0.01

Holdup BI standard deviation: 0.09

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

P value: 0.05

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))