Interaction Details


Details of experiments between P31947 and Q86TI0
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: TBC1D1
Fragment: 232-241
Site: 14-3-3_pSer237
Modification: (SEP)237
Sequence: MRKSFSQPGL
Construct: MRKSF(pSer)QPGL (crude synthetic peptide)
Average holdup BI: 0.6
Holdup BI standard deviation: 0.14
Immobilized partner concentration (10-6M): 113
P value: 2.97
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.12
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: TBC1D1
Fragment: 232-241
Site: 14-3-3_pSer237
Modification: (SEP)237
Sequence: MRKSFSQPGL
Construct: MRKSF(pSer)QPGL (crude synthetic peptide)
Average holdup BI: 0.47
Holdup BI standard deviation: 0.15
Immobilized partner concentration (10-6M): 142
P value: 2.32
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.8
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: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: TBC1D1
Fragment: 232-241
Site: 14-3-3_pSer237
Modification: (SEP)237
Sequence: MRKSFSQPGL
Construct: MRKSF(pSer)QPGL (crude synthetic peptide)
Average holdup BI: 0.56
Holdup BI standard deviation: 0.22
Immobilized partner concentration (10-6M): 111
P value: 2.57
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.07
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: TBC1D1
Fragment: 561-570
Site: 14-3-3_pSer566
Modification: (SEP)566
Sequence: KLLGSSEDLS
Construct: KLLGS(pSer)EDLS (crude synthetic peptide)
Average holdup BI: 0.03
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 113
P value: 2.08
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: TBC1D1
Fragment: 561-570
Site: 14-3-3_pSer566
Modification: (SEP)566
Sequence: KLLGSSEDLS
Construct: KLLGS(pSer)EDLS (crude synthetic peptide)
Average holdup BI: 0.01
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 142
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 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: TBC1D1
Fragment: 561-570
Site: 14-3-3_pSer566
Modification: (SEP)566
Sequence: KLLGSSEDLS
Construct: KLLGS(pSer)EDLS (crude synthetic peptide)
Average holdup BI: 0.03
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 111
P value: 0.78
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: TBC1D1
Fragment: 566-575
Site: 14-3-3_pSer571
Modification: (SEP)571
Sequence: SEDLSSDSES
Construct: SEDLS(pSer)DSES (crude synthetic peptide)
Average holdup BI: -0.46
Holdup BI standard deviation: 1.67
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 8

Protein A

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

Protein B

Protein: TBC1D1
Fragment: 566-575
Site: 14-3-3_pSer571
Modification: (SEP)571
Sequence: SEDLSSDSES
Construct: SEDLS(pSer)DSES (crude synthetic peptide)
Average holdup BI: 0.06
Holdup BI standard deviation: 1.38
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 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: TBC1D1
Fragment: 566-575
Site: 14-3-3_pSer571
Modification: (SEP)571
Sequence: SEDLSSDSES
Construct: SEDLS(pSer)DSES (crude synthetic peptide)
Average holdup BI: -0.01
Holdup BI standard deviation: 1.36
Immobilized partner concentration (10-6M): 111
P value: 0
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: TBC1D1
Fragment: 622-631
Site: 14-3-3_pSer627
Modification: (SEP)627
Sequence: LMRYHSVSTE
Construct: LMRYH(pSer)VSTE (crude synthetic peptide)
Average holdup BI: 0.17
Holdup BI standard deviation: 0.21
Immobilized partner concentration (10-6M): 113
P value: 0.73
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 11

Protein A

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

Protein B

Protein: TBC1D1
Fragment: 622-631
Site: 14-3-3_pSer627
Modification: (SEP)627
Sequence: LMRYHSVSTE
Construct: LMRYH(pSer)VSTE (crude synthetic peptide)
Average holdup BI: -0.02
Holdup BI standard deviation: 0.1
Immobilized partner concentration (10-6M): 142
P value: 0.14
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: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: TBC1D1
Fragment: 622-631
Site: 14-3-3_pSer627
Modification: (SEP)627
Sequence: LMRYHSVSTE
Construct: LMRYH(pSer)VSTE (crude synthetic peptide)
Average holdup BI: -0.15
Holdup BI standard deviation: 0.42
Immobilized partner concentration (10-6M): 111
P value: 0.32
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: TBC1D1
Fragment: 591-600
Site: 14-3-3_pThr596
Modification: (TPO)596
Sequence: RRRANTLSHF
Construct: RRRAN(pThr)LSHF (crude synthetic peptide)
Average holdup BI: -0.02
Holdup BI standard deviation: 0.64
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 14

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: TBC1D1
Fragment: 591-600
Site: 14-3-3_pThr596
Modification: (TPO)596
Sequence: RRRANTLSHF
Construct: RRRAN(pThr)LSHF (crude synthetic peptide)
Average holdup BI: 0.15
Holdup BI standard deviation: 0.23
Immobilized partner concentration (10-6M): 111
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))