Details of experiments between Q04917 and Q07002
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: CDK18
Protein B fragment: 127-136
Protein B site: 14-3-3_pSer132
Protein B modification: (SEP)132
Protein B sequence: MSRRASLSDI
Protein B construct: MSRRA(pSer)LSDI (crude synthetic peptide)
Average holdup BI: 0.62
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): 6.82
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: 4.1
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: CDK18
Protein B fragment: 152-161
Protein B site: 14-3-3_pThr157
Protein B modification: (TPO)157
Protein B sequence: EGTYATVFKG
Protein B construct: EGTYA(pThr)VFKG (crude synthetic peptide)
Average holdup BI: 0.56
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): 6.03
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
Measured pKd: 3.99
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: CDK18
Protein B fragment: 93-102
Protein B site: 14-3-3_pSer98
Protein B modification: (SEP)98
Protein B sequence: VSKRLSLPMD
Protein B construct: VSKRL(pSer)LPMD (crude synthetic peptide)
Average holdup BI: 0.54
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): 5.03
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
Measured pKd: 3.95
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: CDK18
Protein B fragment: 11-20
Protein B site: 14-3-3_pSer16
Protein B modification: (SEP)16
Protein B sequence: RRFSLSVPRT
Protein B construct: RRFSL(pSer)VPRT (crude synthetic peptide)
Average holdup BI: 0.89
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): 4.9
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: 4.77
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: CDK18
Protein B fragment: 84-93
Protein B site: 14-3-3_pSer89
Protein B modification: (SEP)89
Protein B sequence: NQRRFSMEDV
Protein B construct: NQRRF(pSer)MEDV (crude synthetic peptide)
Average holdup BI: 0.24
Holdup BI standard deviation: 0.13
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
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: 3.38