Interaction Details


Details of experiments between P27348 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: YWHAQ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3tau

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: -0.18
Holdup BI standard deviation: 0.37
Immobilized partner concentration (10-6M): 130
P value: 0.54
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: YWHAQ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3tau

Protein B

Protein: TBC1D4
Fragment: 339-348
Site: 14-3-3_pSer344
Modification: (SEP)344
Sequence: HASAPSHVQP
Construct: HASAP(pSer)HVQP (crude synthetic peptide)
Average holdup BI: -0.07
Holdup BI standard deviation: 0.66
Immobilized partner concentration (10-6M): 130
P value: 0.07
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 3

Protein A

Protein: YWHAQ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3tau

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): 130
P value: 0.27
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: YWHAQ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3tau

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: 0.1
Holdup BI standard deviation: 0.15
Immobilized partner concentration (10-6M): 130
P value: 0.68
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 5

Protein A

Protein: YWHAQ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3tau

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: 0.31
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 130
P value: 4.92
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.54
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: YWHAQ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3tau

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: 0.02
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 130
P value: 0.28
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: YWHAQ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3tau

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: 0.1
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 130
P value: 5.67
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: YWHAQ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3tau

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: 0.06
Immobilized partner concentration (10-6M): 130
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 9

Protein A

Protein: YWHAQ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3tau

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: -0.02
Holdup BI standard deviation: 0.16
Immobilized partner concentration (10-6M): 130
P value: 0.11
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: YWHAQ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3tau

Protein B

Protein: TBC1D4
Fragment: 699-708
Site: 14-3-3_pSer704
Modification: (SEP)704
Sequence: LHTSFSAPSF
Construct: LHTSF(pSer)APSF (crude synthetic peptide)
Measured dissociation constant (10-6M): <10
Immobilized partner concentration (10-6M): 130
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 11

Protein A

Protein: YWHAQ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3tau

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: 0.4
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 130
P value: 7.68
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.71
DOI: 10.1101/2022.12.08.519103
Experimental Details

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