Details of experiments between P63104 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: YWHAZ
Protein A fragment: 1-245
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3zeta
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: 0.7
Holdup BI standard deviation: 0.01
Immobilized partner concentration in holdup experiment (10-6M): 132
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 9.28
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: 4.25
Experiment series 2
Protein A protein: YWHAZ
Protein A fragment: 1-245
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3zeta
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: 0.43
Holdup BI standard deviation: 0.06
Immobilized partner concentration in holdup experiment (10-6M): 132
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 4.82
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: 3.76
Experiment series 3
Protein A protein: YWHAZ
Protein A fragment: 1-245
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3zeta
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: 0.13
Holdup BI standard deviation: 0.07
Immobilized partner concentration in holdup experiment (10-6M): 132
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 2.16
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 4
Protein A protein: YWHAZ
Protein A fragment: 1-245
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3zeta
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: 0.12
Holdup BI standard deviation: 0.16
Immobilized partner concentration in holdup experiment (10-6M): 132
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.72
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: YWHAZ
Protein A fragment: 1-245
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3zeta
Protein B protein: WDR62
Protein B fragment: 1129-1138
Protein B site: 14-3-3_pSer1134
Protein B modification: (SEP)1134
Protein B sequence: MDRGGSQPRA
Protein B construct: MDRGG(pSer)QPRA (crude synthetic peptide)
Average holdup BI: 0.15
Holdup BI standard deviation: 0.26
Immobilized partner concentration in holdup experiment (10-6M): 132
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.53
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: YWHAZ
Protein A fragment: 1-245
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3zeta
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
Holdup BI standard deviation: 0.4
Immobilized partner concentration in holdup experiment (10-6M): 132
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.