Interaction Details

Details of experiments between Q9P0K7 and P31946
Note that the results of all experiments are listed, regardless of the modification states of the fragments.

Experiment series 1

Protein A

Protein: RAI14
Fragment: 288-297
Site: 14-3-3_pSer293
Modification: (SEP)293
Sequence: VSSPRSITST
Construct: VSSPR(pSer)ITST (crude synthetic peptide)

Protein B

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

Average holdup BI: 0

Holdup BI standard deviation: 0.06

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

P value: 0.03

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: RAI14
Fragment: 288-297
Site: 14-3-3_pSer293
Modification: (SEP)293
Sequence: VSSPRSITST
Construct: VSSPR(pSer)ITST (crude synthetic peptide)

Protein B

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

Average holdup BI: 0.05

Holdup BI standard deviation: 0.04

Immobilized partner concentration (10^-6M): 102

P value: 1.47

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: RAI14
Fragment: 291-300
Site: 14-3-3_pSer296
Modification: (SEP)296
Sequence: PRSITSTPLS
Construct: PRSIT(pSer)TPLS (crude synthetic peptide)

Protein B

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

Average holdup BI: 0.4

Holdup BI standard deviation: 0.07

Immobilized partner concentration (10^-6M): 102

P value: 4.43

Experiment method: HU Multiplex

Details of affinity fitting: hyperbolic binding equation

Date of measurement: 2021-10-13

Number of measurements: 5

Measured pK_d: 3.81

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: RAI14
Fragment: 291-300
Site: 14-3-3_pSer296
Modification: (SEP)296
Sequence: PRSITSTPLS
Construct: PRSIT(pSer)TPLS (crude synthetic peptide)

Protein B

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

Average holdup BI: 0.45

Holdup BI standard deviation: 0.03

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

P value: 6.06

Experiment method: HU Multiplex

Details of affinity fitting: hyperbolic binding equation

Date of measurement: 2022-02-17

Number of measurements: 4

Measured pK_d: 3.78

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: RAI14
Fragment: 295-304
Site: 14-3-3_pSer300
Modification: (SEP)300
Sequence: TSTPLSGKES
Construct: TSTPL(pSer)GKES (crude synthetic peptide)

Protein B

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

Average holdup BI: -0.07

Holdup BI standard deviation: 0.03

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

P value: 2.3

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: RAI14
Fragment: 295-304
Site: 14-3-3_pSer300
Modification: (SEP)300
Sequence: TSTPLSGKES
Construct: TSTPL(pSer)GKES (crude synthetic peptide)

Protein B

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

Average holdup BI: -0.01

Holdup BI standard deviation: 0.04

Immobilized partner concentration (10^-6M): 102

P value: 0.28

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: RAI14
Fragment: 313-322
Site: 14-3-3_pSer318
Modification: (SEP)318
Sequence: KAEISSIREN
Construct: KAEIS(pSer)IREN (crude synthetic peptide)

Protein B

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

Average holdup BI: 0.03

Holdup BI standard deviation: 0.06

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

P value: 0.51

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 8

Protein A

Protein: RAI14
Fragment: 313-322
Site: 14-3-3_pSer318
Modification: (SEP)318
Sequence: KAEISSIREN
Construct: KAEIS(pSer)IREN (crude synthetic peptide)

Protein B

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

Average holdup BI: 0.01

Holdup BI standard deviation: 0.05

Immobilized partner concentration (10^-6M): 102

P value: 0.16

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: RAI14
Fragment: 353-362
Site: 14-3-3_pSer358
Modification: (SEP)358
Sequence: QAKVASLTLH
Construct: QAKVA(pSer)LTLH (crude synthetic peptide)

Protein B

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

Average holdup BI: 0.27

Holdup BI standard deviation: 0.06

Immobilized partner concentration (10^-6M): 102

P value: 3.88

Experiment method: HU Multiplex

Details of affinity fitting: hyperbolic binding equation

Date of measurement: 2021-10-13

Number of measurements: 5

Measured pK_d: 3.56

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: RAI14
Fragment: 353-362
Site: 14-3-3_pSer358
Modification: (SEP)358
Sequence: QAKVASLTLH
Construct: QAKVA(pSer)LTLH (crude synthetic peptide)

Protein B

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

Average holdup BI: 0.27

Holdup BI standard deviation: 0.05

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

P value: 4.06

Experiment method: HU Multiplex

Details of affinity fitting: hyperbolic binding equation

Date of measurement: 2022-02-17

Number of measurements: 4

Measured pK_d: 3.44

DOI: 10.1101/2022.12.08.519103

Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))

Experiment series 11

Protein A

Protein: RAI14
Fragment: 414-423
Site: 14-3-3_pSer419
Modification: (SEP)419
Sequence: SVLIHSLGKS
Construct: SVLIH(pSer)LGKS (crude synthetic peptide)

Protein B

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

Average holdup BI: 0.11

Holdup BI standard deviation: 0.06

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

P value: 2.07

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: RAI14
Fragment: 414-423
Site: 14-3-3_pSer419
Modification: (SEP)419
Sequence: SVLIHSLGKS
Construct: SVLIH(pSer)LGKS (crude synthetic peptide)

Protein B

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

Average holdup BI: 0.04

Holdup BI standard deviation: 0.06

Immobilized partner concentration (10^-6M): 102

P value: 0.6

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 13

Protein A

Protein: RAI14
Fragment: 662-671
Site: 14-3-3_pSer667
Modification: (SEP)667
Sequence: YRKRKSLEDV
Construct: YRKRK(pSer)LEDV (crude synthetic peptide)

Protein B

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

Average holdup BI: -0.32

Holdup BI standard deviation: 0.67

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

P value: 0.44

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: RAI14
Fragment: 246-255
Site: 14-3-3_pThr251
Modification: (TPO)251
Sequence: DLKTPTKPKQ
Construct: DLKTP(pThr)KPKQ (crude synthetic peptide)

Protein B

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

Average holdup BI: 0

Holdup BI standard deviation: 0.11

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

P value: 0.02

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: RAI14
Fragment: 246-255
Site: 14-3-3_pThr251
Modification: (TPO)251
Sequence: DLKTPTKPKQ
Construct: DLKTP(pThr)KPKQ (crude synthetic peptide)

Protein B

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

Average holdup BI: -0.06

Holdup BI standard deviation: 0.06

Immobilized partner concentration (10^-6M): 102

P value: 1.18

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 16

Protein A

Protein: RAI14
Fragment: 325-334
Site: 14-3-3_pThr330
Modification: (TPO)330
Sequence: RLSDSTTGAD
Construct: RLSDS(pThr)TGAD (crude synthetic peptide)

Protein B

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

Average holdup BI: -0.07

Holdup BI standard deviation: 0.41

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

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))

Experiment series 17

Protein A

Protein: RAI14
Fragment: 325-334
Site: 14-3-3_pThr330
Modification: (TPO)330
Sequence: RLSDSTTGAD
Construct: RLSDS(pThr)TGAD (crude synthetic peptide)

Protein B

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

Average holdup BI: 0.04

Holdup BI standard deviation: 0.69

Immobilized partner concentration (10^-6M): 102

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 18

Protein A

Protein: RAI14
Fragment: 399-408
Site: 14-3-3_pThr404
Modification: (TPO)404
Sequence: GKPGETSPPD
Construct: GKPGE(pThr)SPPD (crude synthetic peptide)

Protein B

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

Average holdup BI: -0.01

Holdup BI standard deviation: 0.11

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

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 19

Protein A

Protein: RAI14
Fragment: 399-408
Site: 14-3-3_pThr404
Modification: (TPO)404
Sequence: GKPGETSPPD
Construct: GKPGE(pThr)SPPD (crude synthetic peptide)

Protein B

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

Average holdup BI: 0.04

Holdup BI standard deviation: 0.19

Immobilized partner concentration (10^-6M): 102

P value: 0.16

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))