Interaction Details


Details of experiments between P31947 and O60343
Note that the results of all experiments are listed, regardless of the modification states of the fragments.


Experiment series 1

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: TBC1D4
Fragment: 1202-1211
Site: 14-3-3_pSer1207
Modification: (SEP)1207
Sequence: LERANSQLKR
Construct: LERAN(pSer)QLKR (crude synthetic peptide)
Average holdup BI: -0.08
Holdup BI standard deviation: 1.17
Immobilized partner concentration (10-6M): 142
P value: 0.06
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 2

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: TBC1D4
Fragment: 1202-1211
Site: 14-3-3_pSer1207
Modification: (SEP)1207
Sequence: LERANSQLKR
Construct: LERAN(pSer)QLKR (crude synthetic peptide)
Average holdup BI: 0.09
Holdup BI standard deviation: 0.61
Immobilized partner concentration (10-6M): 111
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 3

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: TBC1D4
Fragment: 336-345
Site: 14-3-3_pSer341
Modification: (SEP)341
Sequence: RRRHASAPSH
Construct: RRRHA(pSer)APSH (crude synthetic peptide)
Average holdup BI: 0.08
Holdup BI standard deviation: 0.53
Immobilized partner concentration (10-6M): 142
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 4

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: TBC1D4
Fragment: 480-489
Site: 14-3-3_pSer485
Modification: (SEP)485
Sequence: IQRHLSSLTD
Construct: IQRHL(pSer)SLTD (crude synthetic peptide)
Average holdup BI: -0.03
Holdup BI standard deviation: 0.09
Immobilized partner concentration (10-6M): 113
P value: 0.26
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 5

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: TBC1D4
Fragment: 480-489
Site: 14-3-3_pSer485
Modification: (SEP)485
Sequence: IQRHLSSLTD
Construct: IQRHL(pSer)SLTD (crude synthetic peptide)
Average holdup BI: 0
Holdup BI standard deviation: 0.09
Immobilized partner concentration (10-6M): 142
P value: 0.02
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 6

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: TBC1D4
Fragment: 480-489
Site: 14-3-3_pSer485
Modification: (SEP)485
Sequence: IQRHLSSLTD
Construct: IQRHL(pSer)SLTD (crude synthetic peptide)
Average holdup BI: 0
Holdup BI standard deviation: 0.05
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 7

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: TBC1D4
Fragment: 565-574
Site: 14-3-3_pSer570
Modification: (SEP)570
Sequence: RSLTSSLENI
Construct: RSLTS(pSer)LENI (crude synthetic peptide)
Average holdup BI: 0
Holdup BI standard deviation: 0.09
Immobilized partner concentration (10-6M): 113
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 8

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: TBC1D4
Fragment: 565-574
Site: 14-3-3_pSer570
Modification: (SEP)570
Sequence: RSLTSSLENI
Construct: RSLTS(pSer)LENI (crude synthetic peptide)
Average holdup BI: 0.03
Holdup BI standard deviation: 0.13
Immobilized partner concentration (10-6M): 142
P value: 0.2
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: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: TBC1D4
Fragment: 565-574
Site: 14-3-3_pSer570
Modification: (SEP)570
Sequence: RSLTSSLENI
Construct: RSLTS(pSer)LENI (crude synthetic peptide)
Average holdup BI: 0.06
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 111
P value: 0.85
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: TBC1D4
Fragment: 583-592
Site: 14-3-3_pSer588
Modification: (SEP)588
Sequence: RGRLGSVDSF
Construct: RGRLG(pSer)VDSF (crude synthetic peptide)
Average holdup BI: 0.25
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 142
P value: 3.81
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.38
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: TBC1D4
Fragment: 583-592
Site: 14-3-3_pSer588
Modification: (SEP)588
Sequence: RGRLGSVDSF
Construct: RGRLG(pSer)VDSF (crude synthetic peptide)
Average holdup BI: 0.12
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 113
P value: 1.9
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: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: TBC1D4
Fragment: 583-592
Site: 14-3-3_pSer588
Modification: (SEP)588
Sequence: RGRLGSVDSF
Construct: RGRLG(pSer)VDSF (crude synthetic peptide)
Average holdup BI: 0.2
Holdup BI standard deviation: 0.05
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
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: TBC1D4
Fragment: 586-595
Site: 14-3-3_pSer591
Modification: (SEP)591
Sequence: LGSVDSFERS
Construct: LGSVD(pSer)FERS (crude synthetic peptide)
Average holdup BI: 0.04
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 113
P value: 2.38
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: TBC1D4
Fragment: 586-595
Site: 14-3-3_pSer591
Modification: (SEP)591
Sequence: LGSVDSFERS
Construct: LGSVD(pSer)FERS (crude synthetic peptide)
Average holdup BI: 0
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 142
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 15

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: TBC1D4
Fragment: 586-595
Site: 14-3-3_pSer591
Modification: (SEP)591
Sequence: LGSVDSFERS
Construct: LGSVD(pSer)FERS (crude synthetic peptide)
Average holdup BI: 0.01
Holdup BI standard deviation: 0.05
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 16

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: TBC1D4
Fragment: 592-601
Site: 14-3-3_pSer597
Modification: (SEP)597
Sequence: FERSNSLASE
Construct: FERSN(pSer)LASE (crude synthetic peptide)
Average holdup BI: 0.04
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 113
P value: 1.59
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: TBC1D4
Fragment: 592-601
Site: 14-3-3_pSer597
Modification: (SEP)597
Sequence: FERSNSLASE
Construct: FERSN(pSer)LASE (crude synthetic peptide)
Average holdup BI: 0.09
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 142
P value: 4.94
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: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: TBC1D4
Fragment: 592-601
Site: 14-3-3_pSer597
Modification: (SEP)597
Sequence: FERSNSLASE
Construct: FERSN(pSer)LASE (crude synthetic peptide)
Average holdup BI: 0.03
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 111
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 19

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: TBC1D4
Fragment: 595-604
Site: 14-3-3_pSer600
Modification: (SEP)600
Sequence: SNSLASEKDY
Construct: SNSLA(pSer)EKDY (crude synthetic peptide)
Average holdup BI: -0.01
Holdup BI standard deviation: 0.05
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 20

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: TBC1D4
Fragment: 595-604
Site: 14-3-3_pSer600
Modification: (SEP)600
Sequence: SNSLASEKDY
Construct: SNSLA(pSer)EKDY (crude synthetic peptide)
Average holdup BI: -0.02
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 142
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 21

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: TBC1D4
Fragment: 595-604
Site: 14-3-3_pSer600
Modification: (SEP)600
Sequence: SNSLASEKDY
Construct: SNSLA(pSer)EKDY (crude synthetic peptide)
Average holdup BI: 0.03
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 111
P value: 0.65
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: TBC1D4
Fragment: 639-648
Site: 14-3-3_pSer644
Modification: (SEP)644
Sequence: RAHTFSHPPS
Construct: RAHTF(pSer)HPPS (crude synthetic peptide)
Average holdup BI: 0.04
Holdup BI standard deviation: 0.16
Immobilized partner concentration (10-6M): 113
P value: 0.19
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: TBC1D4
Fragment: 639-648
Site: 14-3-3_pSer644
Modification: (SEP)644
Sequence: RAHTFSHPPS
Construct: RAHTF(pSer)HPPS (crude synthetic peptide)
Average holdup BI: 0
Holdup BI standard deviation: 0.13
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 24

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: TBC1D4
Fragment: 639-648
Site: 14-3-3_pSer644
Modification: (SEP)644
Sequence: RAHTFSHPPS
Construct: RAHTF(pSer)HPPS (crude synthetic peptide)
Average holdup BI: 0.14
Holdup BI standard deviation: 0.17
Immobilized partner concentration (10-6M): 111
P value: 0.79
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: TBC1D4
Fragment: 699-708
Site: 14-3-3_pSer704
Modification: (SEP)704
Sequence: LHTSFSAPSF
Construct: LHTSF(pSer)APSF (crude synthetic peptide)
Average holdup BI: 0.88
Holdup BI standard deviation: 0.09
Immobilized partner concentration (10-6M): 142
P value: 1.84
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.71
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: TBC1D4
Fragment: 699-708
Site: 14-3-3_pSer704
Modification: (SEP)704
Sequence: LHTSFSAPSF
Construct: LHTSF(pSer)APSF (crude synthetic peptide)
Average holdup BI: 0.35
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 113
P value: 2.12
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.68
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: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: TBC1D4
Fragment: 699-708
Site: 14-3-3_pSer704
Modification: (SEP)704
Sequence: LHTSFSAPSF
Construct: LHTSF(pSer)APSF (crude synthetic peptide)
Average holdup BI: 0.97
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 111
P value: 6.1
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 5.43
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: TBC1D4
Fragment: 754-763
Site: 14-3-3_pSer759
Modification: (SEP)759
Sequence: SNESLSVGGT
Construct: SNESL(pSer)VGGT (crude synthetic peptide)
Average holdup BI: 0.17
Holdup BI standard deviation: 0.14
Immobilized partner concentration (10-6M): 113
P value: 0.99
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: TBC1D4
Fragment: 563-572
Site: 14-3-3_pThr568
Modification: (TPO)568
Sequence: AKRSLTSSLE
Construct: AKRSL(pThr)SSLE (crude synthetic peptide)
Average holdup BI: 0.29
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 142
P value: 6.05
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.46
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: TBC1D4
Fragment: 563-572
Site: 14-3-3_pThr568
Modification: (TPO)568
Sequence: AKRSLTSSLE
Construct: AKRSL(pThr)SSLE (crude synthetic peptide)
Average holdup BI: 0.22
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 113
P value: 5.93
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.39
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: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: TBC1D4
Fragment: 563-572
Site: 14-3-3_pThr568
Modification: (TPO)568
Sequence: AKRSLTSSLE
Construct: AKRSL(pThr)SSLE (crude synthetic peptide)
Average holdup BI: 0.28
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 111
P value: 7.83
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.54
DOI: 10.1101/2022.12.08.519103
Experimental Details

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