Details of experiments between Q04917 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: YWHAH
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3eta
Protein B protein: TBC1D4
Protein B fragment: 592-601
Protein B site: 14-3-3_pSer597
Protein B modification: (SEP)597
Protein B sequence: FERSNSLASE
Protein B construct: FERSN(pSer)LASE (crude synthetic peptide)
Average holdup BI: 0.19
Holdup BI standard deviation: 0.02
Immobilized partner concentration in holdup experiment (10-6M): 130
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 7.16
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 5
The affinity was below the detection threshold of the assay.
Experiment series 2
Protein A protein: YWHAH
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3eta
Protein B protein: TBC1D4
Protein B fragment: 563-572
Protein B site: 14-3-3_pThr568
Protein B modification: (TPO)568
Protein B sequence: AKRSLTSSLE
Protein B construct: AKRSL(pThr)SSLE (crude synthetic peptide)
Average holdup BI: 0.55
Holdup BI standard deviation: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 130
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 6.64
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 5
Measured pKd: 3.97
Experiment series 3
Protein A protein: YWHAH
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3eta
Protein B protein: TBC1D4
Protein B fragment: 583-592
Protein B site: 14-3-3_pSer588
Protein B modification: (SEP)588
Protein B sequence: RGRLGSVDSF
Protein B construct: RGRLG(pSer)VDSF (crude synthetic peptide)
Average holdup BI: 0.41
Holdup BI standard deviation: 0.04
Immobilized partner concentration in holdup experiment (10-6M): 130
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 5.62
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 5
Measured pKd: 3.73
Experiment series 4
Protein A protein: YWHAH
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3eta
Protein B protein: TBC1D4
Protein B fragment: 595-604
Protein B site: 14-3-3_pSer600
Protein B modification: (SEP)600
Protein B sequence: SNSLASEKDY
Protein B construct: SNSLA(pSer)EKDY (crude synthetic peptide)
Average holdup BI: -0.06
Holdup BI standard deviation: 0.04
Immobilized partner concentration in holdup experiment (10-6M): 130
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 2.03
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 5
The affinity was below the detection threshold of the assay.
Experiment series 5
Protein A protein: YWHAH
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3eta
Protein B protein: TBC1D4
Protein B fragment: 480-489
Protein B site: 14-3-3_pSer485
Protein B modification: (SEP)485
Protein B sequence: IQRHLSSLTD
Protein B construct: IQRHL(pSer)SLTD (crude synthetic peptide)
Average holdup BI: 0.11
Holdup BI standard deviation: 0.07
Immobilized partner concentration in holdup experiment (10-6M): 130
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 1.87
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 5
The affinity was below the detection threshold of the assay.
Experiment series 6
Protein A protein: YWHAH
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3eta
Protein B protein: TBC1D4
Protein B fragment: 565-574
Protein B site: 14-3-3_pSer570
Protein B modification: (SEP)570
Protein B sequence: RSLTSSLENI
Protein B construct: RSLTS(pSer)LENI (crude synthetic peptide)
Average holdup BI: 0.23
Holdup BI standard deviation: 0.15
Immobilized partner concentration in holdup experiment (10-6M): 130
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 1.67
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 5
Measured pKd: 3.37
Experiment series 7
Protein A protein: YWHAH
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3eta
Protein B protein: TBC1D4
Protein B fragment: 586-595
Protein B site: 14-3-3_pSer591
Protein B modification: (SEP)591
Protein B sequence: LGSVDSFERS
Protein B construct: LGSVD(pSer)FERS (crude synthetic peptide)
Average holdup BI: -0.03
Holdup BI standard deviation: 0.04
Immobilized partner concentration in holdup experiment (10-6M): 130
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.84
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 5
The affinity was below the detection threshold of the assay.
Experiment series 8
Protein A protein: YWHAH
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3eta
Protein B protein: TBC1D4
Protein B fragment: 639-648
Protein B site: 14-3-3_pSer644
Protein B modification: (SEP)644
Protein B sequence: RAHTFSHPPS
Protein B construct: RAHTF(pSer)HPPS (crude synthetic peptide)
Average holdup BI: 0
Holdup BI standard deviation: 0.2
Immobilized partner concentration in holdup experiment (10-6M): 130
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.01
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 5
The affinity was below the detection threshold of the assay.