Interaction Details


Details of experiments between Q86TI0 and P61981
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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.9
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 134
P value: 2.72
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.84
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: TBC1D1
Fragment: 232-241
Site: 14-3-3_pSer237
Modification: (SEP)237
Sequence: MRKSFSQPGL
Construct: MRKSF(pSer)QPGL (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.6
Holdup BI standard deviation: 0.12
Immobilized partner concentration (10-6M): 129
P value: 1.96
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.07
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: TBC1D1
Fragment: 498-507
Site: 14-3-3_pSer503
Modification: (SEP)503
Sequence: NKAKRSLTES
Construct: NKAKR(pSer)LTES (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.06
Holdup BI standard deviation: 1.06
Immobilized partner concentration (10-6M): 134
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 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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.08
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 134
P value: 4.7
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: TBC1D1
Fragment: 561-570
Site: 14-3-3_pSer566
Modification: (SEP)566
Sequence: KLLGSSEDLS
Construct: KLLGS(pSer)EDLS (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.04
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 129
P value: 1.35
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: 566-575
Site: 14-3-3_pSer571
Modification: (SEP)571
Sequence: SEDLSSDSES
Construct: SEDLS(pSer)DSES (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: -0.25
Holdup BI standard deviation: 0.66
Immobilized partner concentration (10-6M): 134
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 7

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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.1
Holdup BI standard deviation: 1.43
Immobilized partner concentration (10-6M): 129
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 8

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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.43
Holdup BI standard deviation: 0.11
Immobilized partner concentration (10-6M): 134
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: 3.76
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: TBC1D1
Fragment: 622-631
Site: 14-3-3_pSer627
Modification: (SEP)627
Sequence: LMRYHSVSTE
Construct: LMRYH(pSer)VSTE (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.25
Holdup BI standard deviation: 0.08
Immobilized partner concentration (10-6M): 129
P value: 3.13
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 10

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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.16
Holdup BI standard deviation: 0.55
Immobilized partner concentration (10-6M): 129
P value: 0.23
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 11

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: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.89
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 115
P value: 4.75
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.85
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: TBC1D1
Fragment: 561-570
Site: 14-3-3_pSer566
Modification: (SEP)566
Sequence: KLLGSSEDLS
Construct: KLLGS(pSer)EDLS (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.04
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 115
P value: 1.01
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))