Interaction Details


Details of experiments between P30305 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: CDC25B
Fragment: 146-155
Site: 14-3-3_pSer151
Modification: (SEP)151
Sequence: IRRFQSMPDG
Construct: IRRFQ(pSer)MPDG (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: 0.93
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 134
P value: 5.8
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.97
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: CDC25B
Fragment: 146-155
Site: 14-3-3_pSer151
Modification: (SEP)151
Sequence: IRRFQSMPVR
Construct: IRRFQ(pSer)MPVR (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: 0.9
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 102
P value: 8.21
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.96
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: CDC25B
Fragment: 146-155
Site: 14-3-3_pSer151
Modification: (SEP)151
Sequence: IRRFQSMPDG
Construct: IRRFQ(pSer)MPDG (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: 0.88
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 102
P value: 8.55
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 4

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: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.79
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 134
P value: 4.62
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.45
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: CDC25B
Fragment: 164-173
Site: 14-3-3_pSer169
Modification: (SEP)169
Sequence: RNITNSQAPD
Construct: RNITN(pSer)QAPD (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: -0.12
Holdup BI standard deviation: 0.05
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
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: 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: 0.05
Immobilized partner concentration (10-6M): 102
P value: 1.01
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: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.23
Holdup BI standard deviation: 0.11
Immobilized partner concentration (10-6M): 102
P value: 2.09
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.47
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: CDC25B
Fragment: 204-213
Site: 14-3-3_pSer209
Modification: (SEP)209
Sequence: KPTHPSSTHA
Construct: KPTHP(pSer)STHA (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: -0.33
Holdup BI standard deviation: 0.5
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 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: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.92
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 102
P value: 12.05
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 5.05
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: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.93
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 134
P value: 7.15
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.98
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: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.2
Holdup BI standard deviation: 0.14
Immobilized partner concentration (10-6M): 134
P value: 1.37
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 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: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.02
Holdup BI standard deviation: 0.18
Immobilized partner concentration (10-6M): 102
P value: 0.08
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 13

Protein A

Protein: CDC25B
Fragment: 318-327
Site: 14-3-3_pSer323
Modification: (SEP)323
Sequence: LFRSPSMPCS
Construct: LFRSP(pSer)MPCS (crude synthetic peptide)

Protein B

Protein: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Measured dissociation constant (10-6M): <10
Immobilized partner concentration (10-6M): 102
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 1
Measured pKd: 5
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: 370-379
Site: 14-3-3_pSer375
Modification: (SEP)375
Sequence: VLRSKSLCHD
Construct: VLRSK(pSer)LCHD (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: 0.09
Holdup BI standard deviation: 0.62
Immobilized partner concentration (10-6M): 102
P value: 0.1
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 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: 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: 0.09
Immobilized partner concentration (10-6M): 134
P value: 0.5
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 16

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: 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: 0.07
Immobilized partner concentration (10-6M): 102
P value: 0.9
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 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: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.92
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 134
P value: 7.92
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.91
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: 558-567
Site: 14-3-3_pSer563
Modification: (SEP)563
Sequence: RLKTRSWAGE
Construct: RLKTR(pSer)WAGE (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: 0.89
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 102
P value: 8.52
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.9
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

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: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.37
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 134
P value: 4.01
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.64
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 20

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: 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: 0.08
Immobilized partner concentration (10-6M): 102
P value: 0.58
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 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: 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): 134
P value: 0.11
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 22

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: 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: 0.03
Immobilized partner concentration (10-6M): 102
P value: 0.24
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 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: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.14
Holdup BI standard deviation: 0.08
Immobilized partner concentration (10-6M): 134
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
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 24

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: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.13
Holdup BI standard deviation: 0.12
Immobilized partner concentration (10-6M): 102
P value: 1.1
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 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: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.22
Holdup BI standard deviation: 0.16
Immobilized partner concentration (10-6M): 102
P value: 1.51
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.43
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: 48-57
Site: 14-3-3_pThr53
Modification: (TPO)53
Sequence: ASSPVTTLTQ
Construct: ASSPV(pThr)TLTQ (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: 0.12
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 134
P value: 2.82
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: 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: 0.3
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 28

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: 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: 0.11
Immobilized partner concentration (10-6M): 102
P value: 0.43
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 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: 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: 0.1
Immobilized partner concentration (10-6M): 134
P value: 2.21
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.44
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 30

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: 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: 0.11
Immobilized partner concentration (10-6M): 102
P value: 0.21
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 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: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.38
Holdup BI standard deviation: 0.09
Immobilized partner concentration (10-6M): 134
P value: 3.3
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.66
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 32

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: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.26
Holdup BI standard deviation: 0.1
Immobilized partner concentration (10-6M): 102
P value: 2.85
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.53
DOI: 10.1101/2022.12.08.519103
Experimental Details

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