Details of experiments between Q04917 and Q9UJ41
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: RABGEF1
Protein B fragment: 101-110
Protein B site: 14-3-3_pSer106
Protein B modification: (SEP)106
Protein B sequence: VKKFFSASSR
Protein B construct: VKKFF(pSer)ASSR (crude synthetic peptide)
Average holdup BI: 0.75
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): 6.6
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: 4.36
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: RABGEF1
Protein B fragment: 127-136
Protein B site: 14-3-3_pSer132
Protein B modification: (SEP)132
Protein B sequence: INRQTSIETD
Protein B construct: INRQT(pSer)IETD (crude synthetic peptide)
Average holdup BI: -0.05
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): 0.75
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 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: RABGEF1
Protein B fragment: 108-117
Protein B site: 14-3-3_pSer113
Protein B modification: (SEP)113
Protein B sequence: SSRVGSKKEI
Protein B construct: SSRVG(pSer)KKEI (crude synthetic peptide)
Average holdup BI: 0.06
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): 0.67
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
The affinity was below the detection threshold of the assay.
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: RABGEF1
Protein B fragment: 372-381
Protein B site: 14-3-3_pSer377
Protein B modification: (SEP)377
Protein B sequence: QSLNLSQEDF
Protein B construct: QSLNL(pSer)QEDF (crude synthetic peptide)
Average holdup BI: 0.01
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): 0.1
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 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: RABGEF1
Protein B fragment: 384-393
Protein B site: 14-3-3_pThr389
Protein B modification: (TPO)389
Protein B sequence: YMSGQTSPRK
Protein B construct: YMSGQ(pThr)SPRK (crude synthetic peptide)
Average holdup BI: 0.07
Holdup BI standard deviation: 0.56
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): 0.09
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.