Details of experiments between Q6P1L5 and P31946
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: FAM117B
Protein A fragment: 215-224
Protein A site: 14-3-3_pSer220
Protein A modification: (SEP)220
Protein A sequence: IRRTSSLDTL
Protein A construct: IRRTS(pSer)LDTL (crude synthetic peptide)
Protein B protein: YWHAB
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.76
Holdup BI standard deviation: 0.01
Immobilized partner concentration in holdup experiment (10-6M): 102
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 8.71
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: 4.48
Experiment series 2
Protein A protein: FAM117B
Protein A fragment: 215-224
Protein A site: 14-3-3_pSer220
Protein A modification: (SEP)220
Protein A sequence: IRRTSSLDTL
Protein A construct: IRRTS(pSer)LDTL (crude synthetic peptide)
Protein B protein: YWHAB
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.75
Holdup BI standard deviation: 0.02
Immobilized partner concentration in holdup experiment (10-6M): 134
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 8.17
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: 4.36
Experiment series 3
Protein A protein: FAM117B
Protein A fragment: 33-42
Protein A site: 14-3-3_pThr38
Protein A modification: (TPO)38
Protein A sequence: QPMRATVPFQ
Protein A construct: QPMRA(pThr)VPFQ (crude synthetic peptide)
Protein B protein: YWHAB
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.8
Holdup BI standard deviation: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 102
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 6.07
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
Measured pKd: 4.6
Experiment series 4
Protein A protein: FAM117B
Protein A fragment: 451-460
Protein A site: 14-3-3_pThr456
Protein A modification: (TPO)456
Protein A sequence: LPKYATSPKP
Protein A construct: LPKYA(pThr)SPKP (crude synthetic peptide)
Protein B protein: YWHAB
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.94
Holdup BI standard deviation: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 134
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 4.86
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: 5.07
Experiment series 5
Protein A protein: FAM117B
Protein A fragment: 33-42
Protein A site: 14-3-3_pThr38
Protein A modification: (TPO)38
Protein A sequence: QPMRATVPFQ
Protein A construct: QPMRA(pThr)VPFQ (crude synthetic peptide)
Protein B protein: YWHAB
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.85
Holdup BI standard deviation: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 134
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 4.59
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: 4.61
Experiment series 6
Protein A protein: FAM117B
Protein A fragment: 451-460
Protein A site: 14-3-3_pThr456
Protein A modification: (TPO)456
Protein A sequence: LPKYATSPKP
Protein A construct: LPKYA(pThr)SPKP (crude synthetic peptide)
Protein B protein: YWHAB
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.93
Holdup BI standard deviation: 0.02
Immobilized partner concentration in holdup experiment (10-6M): 102
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 3.28
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: 5.13
Experiment series 7
Protein A protein: FAM117B
Protein A fragment: 268-277
Protein A site: 14-3-3_pSer273
Protein A modification: (SEP)273
Protein A sequence: HKRSASWGST
Protein A construct: HKRSA(pSer)WGST (crude synthetic peptide)
Protein B protein: YWHAB
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: -0.4
Holdup BI standard deviation: 1.01
Immobilized partner concentration in holdup experiment (10-6M): 134
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.45
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 8
Protein A protein: FAM117B
Protein A fragment: 386-395
Protein A site: 14-3-3_pSer391
Protein A modification: (SEP)391
Protein A sequence: SSSTRSIDTQ
Protein A construct: SSSTR(pSer)IDTQ (crude synthetic peptide)
Protein B protein: YWHAB
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: -0.06
Holdup BI standard deviation: 0.17
Immobilized partner concentration in holdup experiment (10-6M): 134
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: 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 9
Protein A protein: FAM117B
Protein A fragment: 386-395
Protein A site: 14-3-3_pSer391
Protein A modification: (SEP)391
Protein A sequence: SSSTRSIDTQ
Protein A construct: SSSTR(pSer)IDTQ (crude synthetic peptide)
Protein B protein: YWHAB
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.06
Holdup BI standard deviation: 0.41
Immobilized partner concentration in holdup experiment (10-6M): 102
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.12
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
The affinity was below the detection threshold of the assay.
Experiment series 10
Protein A protein: FAM117B
Protein A fragment: 204-213
Protein A site: 14-3-3_pSer209
Protein A modification: (SEP)209
Protein A sequence: TRQPSSSPSS
Protein A construct: TRQPS(pSer)SPSS (crude synthetic peptide)
Protein B protein: YWHAB
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.08
Holdup BI standard deviation: 0.92
Immobilized partner concentration in holdup experiment (10-6M): 102
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.06
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.