Interaction Details

Details of experiments between P31946 and O14639
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: ABLIM1
Fragment: 350-359
Site: 14-3-3_pSer355
Modification: (SEP)355
Sequence: RTSSESIYSR
Construct: RTSSE(pSer)IYSR (crude synthetic peptide)

Average holdup BI: 0.06

Holdup BI standard deviation: 0.09

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

P value: 0.67

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

Protein B

Protein: ABLIM1
Fragment: 350-359
Site: 14-3-3_pSer355
Modification: (SEP)355
Sequence: RTSSESIYSR
Construct: RTSSE(pSer)IYSR (crude synthetic peptide)

Average holdup BI: 0.03

Holdup BI standard deviation: 0.04

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

P value: 0.61

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

Protein B

Protein: ABLIM1
Fragment: 353-362
Site: 14-3-3_pSer358
Modification: (SEP)358
Sequence: SESIYSRPGS
Construct: SESIY(pSer)RPGS (crude synthetic peptide)

Average holdup BI: -0.02

Holdup BI standard deviation: 0.02

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

P value: 1.39

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

Protein B

Protein: ABLIM1
Fragment: 353-362
Site: 14-3-3_pSer358
Modification: (SEP)358
Sequence: SESIYSRPGS
Construct: SESIY(pSer)RPGS (crude synthetic peptide)

Average holdup BI: 0.01

Holdup BI standard deviation: 0.02

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

P value: 0.3

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: ABLIM1
Fragment: 358-367
Site: 14-3-3_pSer363
Modification: (SEP)363
Sequence: SRPGSSIPGS
Construct: SRPGS(pSer)IPGS (crude synthetic peptide)

Average holdup BI: 0.03

Holdup BI standard deviation: 0.06

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

P value: 0.48

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

Protein B

Protein: ABLIM1
Fragment: 358-367
Site: 14-3-3_pSer363
Modification: (SEP)363
Sequence: SRPGSSIPGS
Construct: SRPGS(pSer)IPGS (crude synthetic peptide)

Average holdup BI: 0.06

Holdup BI standard deviation: 0.02

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

P value: 3.71

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

Protein B

Protein: ABLIM1
Fragment: 34-43
Site: 14-3-3_pSer39
Modification: (SEP)39
Sequence: NIRRSSSDFF
Construct: NIRRS(pSer)SDFF (crude synthetic peptide)

Average holdup BI: 0.34

Holdup BI standard deviation: 0.13

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

P value: 2.62

Experiment method: HU Multiplex

Details of affinity fitting: hyperbolic binding equation

Date of measurement: 2022-02-17

Number of measurements: 4

Measured pK_d: 3.59

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: ABLIM1
Fragment: 34-43
Site: 14-3-3_pSer39
Modification: (SEP)39
Sequence: NIRRSSSDFF
Construct: NIRRS(pSer)SDFF (crude synthetic peptide)

Average holdup BI: 0.19

Holdup BI standard deviation: 0.06

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

P value: 3.12

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: ABLIM1
Fragment: 417-426
Site: 14-3-3_pSer422
Modification: (SEP)422
Sequence: KQERQSLGES
Construct: KQERQ(pSer)LGES (crude synthetic peptide)

Average holdup BI: -0.33

Holdup BI standard deviation: 1.04

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

P value: 0.35

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: ABLIM1
Fragment: 430-439
Site: 14-3-3_pSer435
Modification: (SEP)435
Sequence: LSPTPSAEGY
Construct: LSPTP(pSer)AEGY (crude synthetic peptide)

Average holdup BI: -0.12

Holdup BI standard deviation: 0.08

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

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 11

Protein A

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

Protein B

Protein: ABLIM1
Fragment: 430-439
Site: 14-3-3_pSer435
Modification: (SEP)435
Sequence: LSPTPSAEGY
Construct: LSPTP(pSer)AEGY (crude synthetic peptide)

Average holdup BI: -0.06

Holdup BI standard deviation: 0.3

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 12

Protein A

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

Protein B

Protein: ABLIM1
Fragment: 447-456
Site: 14-3-3_pSer452
Modification: (SEP)452
Sequence: IHRSTSQGSI
Construct: IHRST(pSer)QGSI (crude synthetic peptide)

Average holdup BI: 0.04

Holdup BI standard deviation: 0.1

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

P value: 0.37

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: ABLIM1
Fragment: 447-456
Site: 14-3-3_pSer452
Modification: (SEP)452
Sequence: IHRSTSQGSI
Construct: IHRST(pSer)QGSI (crude synthetic peptide)

Average holdup BI: -0.25

Holdup BI standard deviation: 0.68

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

P value: 0.4

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 14

Protein A

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

Protein B

Protein: ABLIM1
Fragment: 457-466
Site: 14-3-3_pSer462
Modification: (SEP)462
Sequence: NSPVYSRHSY
Construct: NSPVY(pSer)RHSY (crude synthetic peptide)

Average holdup BI: 0.04

Holdup BI standard deviation: 0.06

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

P value: 0.76

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

Protein B

Protein: ABLIM1
Fragment: 457-466
Site: 14-3-3_pSer462
Modification: (SEP)462
Sequence: NSPVYSRHSY
Construct: NSPVY(pSer)RHSY (crude synthetic peptide)

Average holdup BI: -0.04

Holdup BI standard deviation: 0.09

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

P value: 0.44

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

Protein B

Protein: ABLIM1
Fragment: 650-659
Site: 14-3-3_pSer655
Modification: (SEP)655
Sequence: SSKTASLPGY
Construct: SSKTA(pSer)LPGY (crude synthetic peptide)

Average holdup BI: 0.8

Holdup BI standard deviation: 0.02

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

P value: 9.31

Experiment method: HU Multiplex

Details of affinity fitting: hyperbolic binding equation

Date of measurement: 2021-10-13

Number of measurements: 5

Measured pK_d: 4.59

DOI: 10.1101/2022.12.08.519103

Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 17

Protein A

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

Protein B

Protein: ABLIM1
Fragment: 650-659
Site: 14-3-3_pSer655
Modification: (SEP)655
Sequence: SSKTASLPGY
Construct: SSKTA(pSer)LPGY (crude synthetic peptide)

Average holdup BI: 0.78

Holdup BI standard deviation: 0.01

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

P value: 9.65

Experiment method: HU Multiplex

Details of affinity fitting: hyperbolic binding equation

Date of measurement: 2022-02-17

Number of measurements: 4

Measured pK_d: 4.42

DOI: 10.1101/2022.12.08.519103

Experimental Details

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

Experiment series 18

Protein A

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

Protein B

Protein: ABLIM1
Fragment: 664-673
Site: 14-3-3_pSer669
Modification: (SEP)669
Sequence: LHRPVSTDFA
Construct: LHRPV(pSer)TDFA (crude synthetic peptide)

Average holdup BI: 0.15

Holdup BI standard deviation: 0.11

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

P value: 1.41

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

Protein B

Protein: ABLIM1
Fragment: 664-673
Site: 14-3-3_pSer669
Modification: (SEP)669
Sequence: LHRPVSTDFA
Construct: LHRPV(pSer)TDFA (crude synthetic peptide)

Average holdup BI: 0.06

Holdup BI standard deviation: 0.07

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

P value: 0.89

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 20

Protein A

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

Protein B

Protein: ABLIM1
Fragment: 701-710
Site: 14-3-3_pSer706
Modification: (SEP)706
Sequence: MDRGVSMPNM
Construct: MDRGV(pSer)MPNM (crude synthetic peptide)

Average holdup BI: 0.96

Holdup BI standard deviation: 0.01

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

P value: 3.56

Experiment method: HU Multiplex

Details of affinity fitting: hyperbolic binding equation

Date of measurement: 2022-02-17

Number of measurements: 4

Measured pK_d: 5.27

DOI: 10.1101/2022.12.08.519103

Experimental Details

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

Experiment series 21

Protein A

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

Protein B

Protein: ABLIM1
Fragment: 701-710
Site: 14-3-3_pSer706
Modification: (SEP)706
Sequence: MDRGVSMPNM
Construct: MDRGV(pSer)MPNM (crude synthetic peptide)

Average holdup BI: 0.93

Holdup BI standard deviation: 0.01

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

P value: 7.02

Experiment method: HU Multiplex

Details of affinity fitting: hyperbolic binding equation

Date of measurement: 2021-10-13

Number of measurements: 5

Measured pK_d: 5.13

DOI: 10.1101/2022.12.08.519103

Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 22

Protein A

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

Protein B

Protein: ABLIM1
Fragment: 366-375
Site: 14-3-3_pThr371
Modification: (TPO)371
Sequence: GSPGHTIYAK
Construct: GSPGH(pThr)IYAK (crude synthetic peptide)

Average holdup BI: -0.07

Holdup BI standard deviation: 0.06

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

P value: 1.42

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 23

Protein A

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

Protein B

Protein: ABLIM1
Fragment: 366-375
Site: 14-3-3_pThr371
Modification: (TPO)371
Sequence: GSPGHTIYAK
Construct: GSPGH(pThr)IYAK (crude synthetic peptide)

Average holdup BI: -0.02

Holdup BI standard deviation: 0.1

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

P value: 0.18

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 24

Protein A

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

Protein B

Protein: ABLIM1
Fragment: 424-433
Site: 14-3-3_pThr429
Modification: (TPO)429
Sequence: GESPRTLSPT
Construct: GESPR(pThr)LSPT (crude synthetic peptide)

Average holdup BI: 0.11

Holdup BI standard deviation: 0.02

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

P value: 3.92

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 25

Protein A

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

Protein B

Protein: ABLIM1
Fragment: 424-433
Site: 14-3-3_pThr429
Modification: (TPO)429
Sequence: GESPRTLSPT
Construct: GESPR(pThr)LSPT (crude synthetic peptide)

Average holdup BI: 0.1

Holdup BI standard deviation: 0.02

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

P value: 4.53

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))