Details of experiments between P63104 and Q5TCZ1
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: SH3PXD2A
Protein B fragment: 415-424
Protein B site: 14-3-3_pSer420
Protein B modification: (SEP)420
Protein B sequence: QRAQISSPNL
Protein B construct: QRAQI(pSer)SPNL (crude synthetic peptide)
Average holdup BI: 0.61
Holdup BI standard deviation: 0.03
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): 7.12
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: 4.07
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: SH3PXD2A
Protein B fragment: 374-383
Protein B site: 14-3-3_pSer379
Protein B modification: (SEP)379
Protein B sequence: AKKEISLPIL
Protein B construct: AKKEI(pSer)LPIL (crude synthetic peptide)
Average holdup BI: 0.72
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): 4.58
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: 4.3
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: SH3PXD2A
Protein B fragment: 251-260
Protein B site: 14-3-3_pThr256
Protein B modification: (TPO)256
Protein B sequence: LDRRWTLGGM
Protein B construct: LDRRW(pThr)LGGM (crude synthetic peptide)
Average holdup BI: 0.9
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): 3.19
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: 4.86
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: SH3PXD2A
Protein B fragment: 1047-1056
Protein B site: 14-3-3_pSer1052
Protein B modification: (SEP)1052
Protein B sequence: PAQRNSIPVS
Protein B construct: PAQRN(pSer)IPVS (crude synthetic peptide)
Average holdup BI: 0.05
Holdup BI standard deviation: 0.03
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): 2.09
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: SH3PXD2A
Protein B fragment: 1012-1021
Protein B site: 14-3-3_pSer1017
Protein B modification: (SEP)1017
Protein B sequence: VRRNSSFSTA
Protein B construct: VRRNS(pSer)FSTA (crude synthetic peptide)
Average holdup BI: 0.26
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): 1.6
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: 3.42
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: SH3PXD2A
Protein B fragment: 513-522
Protein B site: 14-3-3_pThr518
Protein B modification: (TPO)518
Protein B sequence: RTSTLTRPKV
Protein B construct: RTSTL(pThr)RPKV (crude synthetic peptide)
Average holdup BI: 0.05
Holdup BI standard deviation: 0.05
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): 1.1
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 7
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: SH3PXD2A
Protein B fragment: 997-1006
Protein B site: 14-3-3_pSer1002
Protein B modification: (SEP)1002
Protein B sequence: LRRNESLTAT
Protein B construct: LRRNE(pSer)LTAT (crude synthetic peptide)
Average holdup BI: 0.03
Holdup BI standard deviation: 0.12
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): 0.18
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: 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: SH3PXD2A
Protein B fragment: 633-642
Protein B site: 14-3-3_pSer638
Protein B modification: (SEP)638
Protein B sequence: LSARGSSGDS
Protein B construct: LSARG(pSer)SGDS (crude synthetic peptide)
Average holdup BI: 0.01
Holdup BI standard deviation: 0.38
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): 0.02
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.