Details of experiments between P62258 and Q9P2F8
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Protein B protein: SIPA1L2
Protein B fragment: 157-166
Protein B site: 14-3-3_pSer162
Protein B modification: (SEP)162
Protein B sequence: RQRSNSDVTI
Protein B construct: RQRSN(pSer)DVTI (crude synthetic peptide)
Average holdup BI: 0.4
Holdup BI standard deviation: 0.07
Immobilized partner concentration in holdup experiment (10-6M): 116
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 4.44
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 4
Measured pKd: 3.76
Experiment series 2
Protein A protein: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Protein B protein: SIPA1L2
Protein B fragment: 157-166
Protein B site: 14-3-3_pSer162
Protein B modification: (SEP)162
Protein B sequence: RQRSNSDVTI
Protein B construct: RQRSN(pSer)DVTI (crude synthetic peptide)
Average holdup BI: 0.35
Holdup BI standard deviation: 0.07
Immobilized partner concentration in holdup experiment (10-6M): 143
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 4.14
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: 3.57
Experiment series 3
Protein A protein: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Protein B protein: SIPA1L2
Protein B fragment: 1467-1476
Protein B site: 14-3-3_pSer1472
Protein B modification: (SEP)1472
Protein B sequence: GRRKFSFYGN
Protein B construct: GRRKF(pSer)FYGN (crude synthetic peptide)
Average holdup BI: -0.34
Holdup BI standard deviation: 0.24
Immobilized partner concentration in holdup experiment (10-6M): 116
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 1.82
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 4
The affinity was below the detection threshold of the assay.
Experiment series 4
Protein A protein: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Protein B protein: SIPA1L2
Protein B fragment: 1483-1492
Protein B site: 14-3-3_pSer1488
Protein B modification: (SEP)1488
Protein B sequence: LYRTLSDESI
Protein B construct: LYRTL(pSer)DESI (crude synthetic peptide)
Average holdup BI: 0.06
Holdup BI standard deviation: 0.06
Immobilized partner concentration in holdup experiment (10-6M): 143
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 1.03
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
The affinity was below the detection threshold of the assay.
Experiment series 5
Protein A protein: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Protein B protein: SIPA1L2
Protein B fragment: 1483-1492
Protein B site: 14-3-3_pSer1488
Protein B modification: (SEP)1488
Protein B sequence: LYRTLSDESI
Protein B construct: LYRTL(pSer)DESI (crude synthetic peptide)
Average holdup BI: 0.06
Holdup BI standard deviation: 0.15
Immobilized partner concentration in holdup experiment (10-6M): 116
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.31
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 4
The affinity was below the detection threshold of the assay.
Experiment series 6
Protein A protein: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Protein B protein: SIPA1L2
Protein B fragment: 1467-1476
Protein B site: 14-3-3_pSer1472
Protein B modification: (SEP)1472
Protein B sequence: GRRKFSFYGN
Protein B construct: GRRKF(pSer)FYGN (crude synthetic peptide)
Average holdup BI: 0
Holdup BI standard deviation: 0.13
Immobilized partner concentration in holdup experiment (10-6M): 143
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.