Interaction Details


Details of experiments between P30305 and P31947
Note that the results of all experiments are listed, regardless of the modification states of the fragments.


Experiment series 1

Protein A

Protein: CDC25B
Fragment: 146-155
Site: 14-3-3_pSer151
Modification: (SEP)151
Sequence: IRRFQSMPVR
Construct: IRRFQ(pSer)MPVR (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.91
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 142
P value: 8.1
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.84
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: CDC25B
Fragment: 146-155
Site: 14-3-3_pSer151
Modification: (SEP)151
Sequence: IRRFQSMPVR
Construct: IRRFQ(pSer)MPVR (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.79
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 113
P value: 4.67
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.53
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: CDC25B
Fragment: 146-155
Site: 14-3-3_pSer151
Modification: (SEP)151
Sequence: IRRFQSMPDG
Construct: IRRFQ(pSer)MPDG (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.65
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 113
P value: 4.94
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.21
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: CDC25B
Fragment: 146-155
Site: 14-3-3_pSer151
Modification: (SEP)151
Sequence: IRRFQSMPDG
Construct: IRRFQ(pSer)MPDG (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.7
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 142
P value: 7.71
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.2
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: CDC25B
Fragment: 164-173
Site: 14-3-3_pSer169
Modification: (SEP)169
Sequence: RNITNSQAPD
Construct: RNITN(pSer)QAPD (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: -0.02
Holdup BI standard deviation: 0.1
Immobilized partner concentration (10-6M): 113
P value: 0.17
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: CDC25B
Fragment: 164-173
Site: 14-3-3_pSer169
Modification: (SEP)169
Sequence: RNITNSQAPD
Construct: RNITN(pSer)QAPD (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 142
P value: 0
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: CDC25B
Fragment: 204-213
Site: 14-3-3_pSer209
Modification: (SEP)209
Sequence: KPTHPSSTHA
Construct: KPTHP(pSer)STHA (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: -0.2
Holdup BI standard deviation: 0.64
Immobilized partner concentration (10-6M): 113
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 8

Protein A

Protein: CDC25B
Fragment: 204-213
Site: 14-3-3_pSer209
Modification: (SEP)209
Sequence: KPTHPSSTHA
Construct: KPTHP(pSer)STHA (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.07
Holdup BI standard deviation: 0.17
Immobilized partner concentration (10-6M): 142
P value: 0.34
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: CDC25B
Fragment: 225-234
Site: 14-3-3_pSer230
Modification: (SEP)230
Sequence: AQRPSSAPDL
Construct: AQRPS(pSer)APDL (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.8
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 142
P value: 10.48
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.45
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: CDC25B
Fragment: 225-234
Site: 14-3-3_pSer230
Modification: (SEP)230
Sequence: AQRPSSAPDL
Construct: AQRPS(pSer)APDL (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.75
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 113
P value: 6.51
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.41
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: CDC25B
Fragment: 286-295
Site: 14-3-3_pSer291
Modification: (SEP)291
Sequence: PPGMESLISA
Construct: PPGME(pSer)LISA (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.15
Holdup BI standard deviation: 0.13
Immobilized partner concentration (10-6M): 113
P value: 1.02
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 12

Protein A

Protein: CDC25B
Fragment: 286-295
Site: 14-3-3_pSer291
Modification: (SEP)291
Sequence: PPGMESLISA
Construct: PPGME(pSer)LISA (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: -0.16
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 142
P value: 2.99
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: CDC25B
Fragment: 370-379
Site: 14-3-3_pSer375
Modification: (SEP)375
Sequence: VLRSKSLCHD
Construct: VLRSK(pSer)LCHD (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: -0.38
Holdup BI standard deviation: 0.53
Immobilized partner concentration (10-6M): 142
P value: 0.77
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: CDC25B
Fragment: 460-469
Site: 14-3-3_pSer465
Modification: (SEP)465
Sequence: ERDAESFLLK
Construct: ERDAE(pSer)FLLK (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: -0.07
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 113
P value: 1.55
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: CDC25B
Fragment: 460-469
Site: 14-3-3_pSer465
Modification: (SEP)465
Sequence: ERDAESFLLK
Construct: ERDAE(pSer)FLLK (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0
Holdup BI standard deviation: 0.08
Immobilized partner concentration (10-6M): 142
P value: 0.03
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: CDC25B
Fragment: 558-567
Site: 14-3-3_pSer563
Modification: (SEP)563
Sequence: RLKTRSWAGE
Construct: RLKTR(pSer)WAGE (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.72
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 142
P value: 7.43
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.25
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: CDC25B
Fragment: 558-567
Site: 14-3-3_pSer563
Modification: (SEP)563
Sequence: RLKTRSWAGE
Construct: RLKTR(pSer)WAGE (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.63
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 113
P value: 7.05
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.17
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: CDC25B
Fragment: 122-131
Site: 14-3-3_pThr127
Modification: (TPO)127
Sequence: HMAEQTFEQA
Construct: HMAEQ(pThr)FEQA (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.21
Holdup BI standard deviation: 0.12
Immobilized partner concentration (10-6M): 113
P value: 1.73
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.37
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: CDC25B
Fragment: 122-131
Site: 14-3-3_pThr127
Modification: (TPO)127
Sequence: HMAEQTFEQA
Construct: HMAEQ(pThr)FEQA (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.07
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 142
P value: 1.04
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 20

Protein A

Protein: CDC25B
Fragment: 162-171
Site: 14-3-3_pThr167
Modification: (TPO)167
Sequence: VLRNITNSQA
Construct: VLRNI(pThr)NSQA (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 113
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 21

Protein A

Protein: CDC25B
Fragment: 162-171
Site: 14-3-3_pThr167
Modification: (TPO)167
Sequence: VLRNITNSQA
Construct: VLRNI(pThr)NSQA (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: -0.01
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 142
P value: 0.34
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 22

Protein A

Protein: CDC25B
Fragment: 399-408
Site: 14-3-3_pThr404
Modification: (TPO)404
Sequence: AFLLQTVDGK
Construct: AFLLQ(pThr)VDGK (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.04
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 113
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 23

Protein A

Protein: CDC25B
Fragment: 399-408
Site: 14-3-3_pThr404
Modification: (TPO)404
Sequence: AFLLQTVDGK
Construct: AFLLQ(pThr)VDGK (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.02
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 142
P value: 0.26
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: CDC25B
Fragment: 48-57
Site: 14-3-3_pThr53
Modification: (TPO)53
Sequence: ASSPVTTLTQ
Construct: ASSPV(pThr)TLTQ (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.07
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 113
P value: 2.26
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: CDC25B
Fragment: 48-57
Site: 14-3-3_pThr53
Modification: (TPO)53
Sequence: ASSPVTTLTQ
Construct: ASSPV(pThr)TLTQ (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.18
Holdup BI standard deviation: 0.15
Immobilized partner concentration (10-6M): 142
P value: 1.29
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 26

Protein A

Protein: CDC25B
Fragment: 49-58
Site: 14-3-3_pThr54
Modification: (TPO)54
Sequence: SSPVTTLTQT
Construct: SSPVT(pThr)LTQT (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.04
Holdup BI standard deviation: 0.14
Immobilized partner concentration (10-6M): 113
P value: 0.24
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 27

Protein A

Protein: CDC25B
Fragment: 49-58
Site: 14-3-3_pThr54
Modification: (TPO)54
Sequence: SSPVTTLTQT
Construct: SSPVT(pThr)LTQT (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.01
Holdup BI standard deviation: 0.1
Immobilized partner concentration (10-6M): 142
P value: 0.05
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 28

Protein A

Protein: CDC25B
Fragment: 51-60
Site: 14-3-3_pThr56
Modification: (TPO)56
Sequence: PVTTLTQTMH
Construct: PVTTL(pThr)QTMH (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.12
Holdup BI standard deviation: 0.15
Immobilized partner concentration (10-6M): 113
P value: 0.74
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 29

Protein A

Protein: CDC25B
Fragment: 51-60
Site: 14-3-3_pThr56
Modification: (TPO)56
Sequence: PVTTLTQTMH
Construct: PVTTL(pThr)QTMH (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: -0.07
Holdup BI standard deviation: 0.11
Immobilized partner concentration (10-6M): 142
P value: 0.68
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 30

Protein A

Protein: CDC25B
Fragment: 53-62
Site: 14-3-3_pThr58
Modification: (TPO)58
Sequence: TTLTQTMHDL
Construct: TTLTQ(pThr)MHDL (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.15
Holdup BI standard deviation: 0.17
Immobilized partner concentration (10-6M): 113
P value: 0.88
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 31

Protein A

Protein: CDC25B
Fragment: 53-62
Site: 14-3-3_pThr58
Modification: (TPO)58
Sequence: TTLTQTMHDL
Construct: TTLTQ(pThr)MHDL (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.1
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 142
P value: 2.58
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 32

Protein A

Protein: CDC25B
Fragment: 146-155
Site: 14-3-3_pSer151
Modification: (SEP)151
Sequence: IRRFQSMPVR
Construct: IRRFQ(pSer)MPVR (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.97
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 111
P value: 3.28
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 5.44
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 33

Protein A

Protein: CDC25B
Fragment: 146-155
Site: 14-3-3_pSer151
Modification: (SEP)151
Sequence: IRRFQSMPDG
Construct: IRRFQ(pSer)MPDG (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.06
Holdup BI standard deviation: 0.1
Immobilized partner concentration (10-6M): 111
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 34

Protein A

Protein: CDC25B
Fragment: 164-173
Site: 14-3-3_pSer169
Modification: (SEP)169
Sequence: RNITNSQAPD
Construct: RNITN(pSer)QAPD (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.01
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 111
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 35

Protein A

Protein: CDC25B
Fragment: 204-213
Site: 14-3-3_pSer209
Modification: (SEP)209
Sequence: KPTHPSSTHA
Construct: KPTHP(pSer)STHA (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.08
Holdup BI standard deviation: 0.27
Immobilized partner concentration (10-6M): 111
P value: 0.23
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 36

Protein A

Protein: CDC25B
Fragment: 225-234
Site: 14-3-3_pSer230
Modification: (SEP)230
Sequence: AQRPSSAPDL
Construct: AQRPS(pSer)APDL (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.69
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 111
P value: 10.54
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.3
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 37

Protein A

Protein: CDC25B
Fragment: 286-295
Site: 14-3-3_pSer291
Modification: (SEP)291
Sequence: PPGMESLISA
Construct: PPGME(pSer)LISA (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: -0.14
Holdup BI standard deviation: 0.27
Immobilized partner concentration (10-6M): 111
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 38

Protein A

Protein: CDC25B
Fragment: 370-379
Site: 14-3-3_pSer375
Modification: (SEP)375
Sequence: VLRSKSLCHD
Construct: VLRSK(pSer)LCHD (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.06
Holdup BI standard deviation: 0.73
Immobilized partner concentration (10-6M): 111
P value: 0.06
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 39

Protein A

Protein: CDC25B
Fragment: 460-469
Site: 14-3-3_pSer465
Modification: (SEP)465
Sequence: ERDAESFLLK
Construct: ERDAE(pSer)FLLK (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.01
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 111
P value: 0.12
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 40

Protein A

Protein: CDC25B
Fragment: 558-567
Site: 14-3-3_pSer563
Modification: (SEP)563
Sequence: RLKTRSWAGE
Construct: RLKTR(pSer)WAGE (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.62
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 111
P value: 4.22
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.17
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 41

Protein A

Protein: CDC25B
Fragment: 122-131
Site: 14-3-3_pThr127
Modification: (TPO)127
Sequence: HMAEQTFEQA
Construct: HMAEQ(pThr)FEQA (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: -0.11
Holdup BI standard deviation: 0.28
Immobilized partner concentration (10-6M): 111
P value: 0.36
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 42

Protein A

Protein: CDC25B
Fragment: 162-171
Site: 14-3-3_pThr167
Modification: (TPO)167
Sequence: VLRNITNSQA
Construct: VLRNI(pThr)NSQA (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 111
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 43

Protein A

Protein: CDC25B
Fragment: 399-408
Site: 14-3-3_pThr404
Modification: (TPO)404
Sequence: AFLLQTVDGK
Construct: AFLLQ(pThr)VDGK (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.06
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 111
P value: 0.91
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 44

Protein A

Protein: CDC25B
Fragment: 48-57
Site: 14-3-3_pThr53
Modification: (TPO)53
Sequence: ASSPVTTLTQ
Construct: ASSPV(pThr)TLTQ (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.01
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 111
P value: 0.24
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 45

Protein A

Protein: CDC25B
Fragment: 49-58
Site: 14-3-3_pThr54
Modification: (TPO)54
Sequence: SSPVTTLTQT
Construct: SSPVT(pThr)LTQT (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: -0.17
Holdup BI standard deviation: 0.29
Immobilized partner concentration (10-6M): 111
P value: 0.63
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 46

Protein A

Protein: CDC25B
Fragment: 51-60
Site: 14-3-3_pThr56
Modification: (TPO)56
Sequence: PVTTLTQTMH
Construct: PVTTL(pThr)QTMH (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: -0.19
Holdup BI standard deviation: 0.25
Immobilized partner concentration (10-6M): 111
P value: 0.8
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 47

Protein A

Protein: CDC25B
Fragment: 53-62
Site: 14-3-3_pThr58
Modification: (TPO)58
Sequence: TTLTQTMHDL
Construct: TTLTQ(pThr)MHDL (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.01
Holdup BI standard deviation: 0.28
Immobilized partner concentration (10-6M): 111
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))