Interaction Details


Details of experiments between Q86TI0 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: TBC1D1
Fragment: 232-241
Site: 14-3-3_pSer237
Modification: (SEP)237
Sequence: MRKSFSQPGL
Construct: MRKSF(pSer)QPGL (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: 0.65
Holdup BI standard deviation: 0.12
Immobilized partner concentration (10-6M): 102
P value: 2.55
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.25
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: TBC1D1
Fragment: 232-241
Site: 14-3-3_pSer237
Modification: (SEP)237
Sequence: MRKSFSQPGL
Construct: MRKSF(pSer)QPGL (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): <100
Immobilized partner concentration (10-6M): 134
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 1
Measured pKd: 4
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: TBC1D1
Fragment: 561-570
Site: 14-3-3_pSer566
Modification: (SEP)566
Sequence: KLLGSSEDLS
Construct: KLLGS(pSer)EDLS (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: 0.02
Immobilized partner concentration (10-6M): 134
P value: 1.09
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 4

Protein A

Protein: TBC1D1
Fragment: 561-570
Site: 14-3-3_pSer566
Modification: (SEP)566
Sequence: KLLGSSEDLS
Construct: KLLGS(pSer)EDLS (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: 0.08
Holdup BI standard deviation: 0.12
Immobilized partner concentration (10-6M): 102
P value: 0.65
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: TBC1D1
Fragment: 566-575
Site: 14-3-3_pSer571
Modification: (SEP)571
Sequence: SEDLSSDSES
Construct: SEDLS(pSer)DSES (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: 1.48
Immobilized partner concentration (10-6M): 102
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: TBC1D1
Fragment: 622-631
Site: 14-3-3_pSer627
Modification: (SEP)627
Sequence: LMRYHSVSTE
Construct: LMRYH(pSer)VSTE (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: 0.47
Holdup BI standard deviation: 0.1
Immobilized partner concentration (10-6M): 134
P value: 3.2
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.81
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: TBC1D1
Fragment: 622-631
Site: 14-3-3_pSer627
Modification: (SEP)627
Sequence: LMRYHSVSTE
Construct: LMRYH(pSer)VSTE (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: 0.19
Holdup BI standard deviation: 0.08
Immobilized partner concentration (10-6M): 102
P value: 2.68
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 8

Protein A

Protein: TBC1D1
Fragment: 591-600
Site: 14-3-3_pThr596
Modification: (TPO)596
Sequence: RRRANTLSHF
Construct: RRRAN(pThr)LSHF (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: -0.16
Holdup BI standard deviation: 0.68
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))