Details of experiments between P62258 and Q6P1L5
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: FAM117B
Protein B fragment: 451-460
Protein B site: 14-3-3_pThr456
Protein B modification: (TPO)456
Protein B sequence: LPKYATSPKP
Protein B construct: LPKYA(pThr)SPKP (crude synthetic peptide)
Average holdup BI: 0.88
Holdup BI standard deviation: 0.01
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): 8.07
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: 4.8
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: FAM117B
Protein B fragment: 215-224
Protein B site: 14-3-3_pSer220
Protein B modification: (SEP)220
Protein B sequence: IRRTSSLDTL
Protein B construct: IRRTS(pSer)LDTL (crude synthetic peptide)
Average holdup BI: 0.52
Holdup BI standard deviation: 0.03
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): 7.04
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: 3.87
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: FAM117B
Protein B fragment: 33-42
Protein B site: 14-3-3_pThr38
Protein B modification: (TPO)38
Protein B sequence: QPMRATVPFQ
Protein B construct: QPMRA(pThr)VPFQ (crude synthetic peptide)
Average holdup BI: 0.76
Holdup BI standard deviation: 0.05
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): 5.78
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: 4.34
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: FAM117B
Protein B fragment: 215-224
Protein B site: 14-3-3_pSer220
Protein B modification: (SEP)220
Protein B sequence: IRRTSSLDTL
Protein B construct: IRRTS(pSer)LDTL (crude synthetic peptide)
Average holdup BI: 0.43
Holdup BI standard deviation: 0.05
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): 5.64
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: 3.82
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: FAM117B
Protein B fragment: 451-460
Protein B site: 14-3-3_pThr456
Protein B modification: (TPO)456
Protein B sequence: LPKYATSPKP
Protein B construct: LPKYA(pThr)SPKP (crude synthetic peptide)
Average holdup BI: 0.93
Holdup BI standard deviation: 0.02
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): 5.23
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: 4.95
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: FAM117B
Protein B fragment: 33-42
Protein B site: 14-3-3_pThr38
Protein B modification: (TPO)38
Protein B sequence: QPMRATVPFQ
Protein B construct: QPMRA(pThr)VPFQ (crude synthetic peptide)
Average holdup BI: 0.76
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): 4.2
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: 4.44
Experiment series 7
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: FAM117B
Protein B fragment: 386-395
Protein B site: 14-3-3_pSer391
Protein B modification: (SEP)391
Protein B sequence: SSSTRSIDTQ
Protein B construct: SSSTR(pSer)IDTQ (crude synthetic peptide)
Average holdup BI: 0.12
Holdup BI standard deviation: 0.2
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): 0.54
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: 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: FAM117B
Protein B fragment: 386-395
Protein B site: 14-3-3_pSer391
Protein B modification: (SEP)391
Protein B sequence: SSSTRSIDTQ
Protein B construct: SSSTR(pSer)IDTQ (crude synthetic peptide)
Average holdup BI: -0.09
Holdup BI standard deviation: 0.38
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): 0.22
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: 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: FAM117B
Protein B fragment: 268-277
Protein B site: 14-3-3_pSer273
Protein B modification: (SEP)273
Protein B sequence: HKRSASWGST
Protein B construct: HKRSA(pSer)WGST (crude synthetic peptide)
Average holdup BI: -0.21
Holdup BI standard deviation: 0.94
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): 0.2
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 10
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: FAM117B
Protein B fragment: 204-213
Protein B site: 14-3-3_pSer209
Protein B modification: (SEP)209
Protein B sequence: TRQPSSSPSS
Protein B construct: TRQPS(pSer)SPSS (crude synthetic peptide)
Average holdup BI: 0.11
Holdup BI standard deviation: 0.63
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): 0.12
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.