Details of experiments between P27348 and O43379
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: YWHAQ
Protein A fragment: 1-245
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3tau
Protein B protein: WDR62
Protein B fragment: 1223-1232
Protein B site: 14-3-3_pSer1228
Protein B modification: (SEP)1228
Protein B sequence: ISRSISLGDS
Protein B construct: ISRSI(pSer)LGDS (crude synthetic peptide)
Average holdup BI: 0.68
Holdup BI standard deviation: 0.03
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): 8.31
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: 4.21
Experiment series 2
Protein A protein: YWHAQ
Protein A fragment: 1-245
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3tau
Protein B protein: WDR62
Protein B fragment: 1244-1253
Protein B site: 14-3-3_pSer1249
Protein B modification: (SEP)1249
Protein B sequence: LRRPSSVGEL
Protein B construct: LRRPS(pSer)VGEL (crude synthetic peptide)
Average holdup BI: 0.4
Holdup BI standard deviation: 0.06
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): 4.67
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: 3.7
Experiment series 3
Protein A protein: YWHAQ
Protein A fragment: 1-245
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3tau
Protein B protein: WDR62
Protein B fragment: 44-53
Protein B site: 14-3-3_pSer49
Protein B modification: (SEP)49
Protein B sequence: RRTRLSTASE
Protein B construct: RRTRL(pSer)TASE (crude synthetic peptide)
Average holdup BI: 0.22
Holdup BI standard deviation: 0.12
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): 1.85
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: 3.33
Experiment series 4
Protein A protein: YWHAQ
Protein A fragment: 1-245
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3tau
Protein B protein: WDR62
Protein B fragment: 892-901
Protein B site: 14-3-3_pSer897
Protein B modification: (SEP)897
Protein B sequence: ESLENSILDS
Protein B construct: ESLEN(pSer)ILDS (crude synthetic peptide)
Average holdup BI: -0.27
Holdup BI standard deviation: 0.28
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): 1.49
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: YWHAQ
Protein A fragment: 1-245
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3tau
Protein B protein: WDR62
Protein B fragment: 1337-1346
Protein B site: 14-3-3_pSer1342
Protein B modification: (SEP)1342
Protein B sequence: SAFRPSLPAP
Protein B construct: SAFRP(pSer)LPAP (crude synthetic peptide)
Average holdup BI: 0.09
Holdup BI standard deviation: 0.1
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): 0.98
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.