Details of experiments between Q86SQ0 and Q04917
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: PHLDB2
Protein A fragment: 195-204
Protein A site: 14-3-3_pSer200
Protein A modification: (SEP)200
Protein A sequence: RSGAASMPSS
Protein A construct: RSGAA(pSer)MPSS (crude synthetic peptide)
Protein B protein: YWHAH
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3eta
Average holdup BI: 0.83
Holdup BI standard deviation: 0.04
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): 8.56
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: 4.59
Experiment series 2
Protein A protein: PHLDB2
Protein A fragment: 272-281
Protein A site: 14-3-3_pSer277
Protein A modification: (SEP)277
Protein A sequence: LPPRNSLGNS
Protein A construct: LPPRN(pSer)LGNS (crude synthetic peptide)
Protein B protein: YWHAH
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3eta
Average holdup BI: 0.13
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): 3.91
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: PHLDB2
Protein A fragment: 893-902
Protein A site: 14-3-3_pThr898
Protein A modification: (TPO)898
Protein A sequence: LYLSDTLPRK
Protein A construct: LYLSD(pThr)LPRK (crude synthetic peptide)
Protein B protein: YWHAH
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3eta
Average holdup BI: 0.28
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): 2.46
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: 3.47
Experiment series 4
Protein A protein: PHLDB2
Protein A fragment: 410-419
Protein A site: 14-3-3_pSer415
Protein A modification: (SEP)415
Protein A sequence: RERKSSISSI
Protein A construct: RERKS(pSer)ISSI (crude synthetic peptide)
Protein B protein: YWHAH
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3eta
Average holdup BI: 0.11
Holdup BI standard deviation: 0.14
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): 0.7
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: PHLDB2
Protein A fragment: 237-246
Protein A site: 14-3-3_pSer242
Protein A modification: (SEP)242
Protein A sequence: RTRKYSSSSL
Protein A construct: RTRKY(pSer)SSSL (crude synthetic peptide)
Protein B protein: YWHAH
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3eta
Average holdup BI: 0.17
Holdup BI standard deviation: 0.23
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): 0.57
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: PHLDB2
Protein A fragment: 484-493
Protein A site: 14-3-3_pSer489
Protein A modification: (SEP)489
Protein A sequence: EKLQLSDEES
Protein A construct: EKLQL(pSer)DEES (crude synthetic peptide)
Protein B protein: YWHAH
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3eta
Average holdup BI: -0.07
Holdup BI standard deviation: 0.17
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): 0.42
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: PHLDB2
Protein A fragment: 463-472
Protein A site: 14-3-3_pSer468
Protein A modification: (SEP)468
Protein A sequence: PDSRLSTGTT
Protein A construct: PDSRL(pSer)TGTT (crude synthetic peptide)
Protein B protein: YWHAH
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3eta
Average holdup BI: 0.01
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): 0.3
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: PHLDB2
Protein A fragment: 1123-1132
Protein A site: 14-3-3_pThr1128
Protein A modification: (TPO)1128
Protein A sequence: RSHVETAGHN
Protein A construct: RSHVE(pThr)AGHN (crude synthetic peptide)
Protein B protein: YWHAH
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3eta
Average holdup BI: 0.18
Holdup BI standard deviation: 0.48
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): 0.24
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 9
Protein A protein: PHLDB2
Protein A fragment: 207-216
Protein A site: 14-3-3_pSer212
Protein A modification: (SEP)212
Protein A sequence: QARKMSIQDS
Protein A construct: QARKM(pSer)IQDS (crude synthetic peptide)
Protein B protein: YWHAH
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3eta
Average holdup BI: 0.01
Holdup BI standard deviation: 0.14
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): 0.07
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.