Details of experiments between Q9BZF3 and P31947
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: OSBPL6
Protein A fragment: 285-294
Protein A site: 14-3-3_pSer290
Protein A modification: (SEP)290
Protein A sequence: LQRTQSAPNF
Protein A construct: LQRTQ(pSer)APNF (crude synthetic peptide)
Protein B protein: SFN
Protein B fragment: 1-248
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3sigma
Measured dissociation constant (10-6M): <1
Immobilized partner concentration in holdup experiment (10-6M): 113
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: 1
Measured pKd: 6
Experiment series 2
Protein A protein: OSBPL6
Protein A fragment: 285-294
Protein A site: 14-3-3_pSer290
Protein A modification: (SEP)290
Protein A sequence: LQRTQSAPNF
Protein A construct: LQRTQ(pSer)APNF (crude synthetic peptide)
Protein B protein: SFN
Protein B fragment: 1-248
Protein B site: 14-3-3
Protein B modification: 100_uM_FCA
Protein B construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.67
Holdup BI standard deviation: 0.02
Immobilized partner concentration in holdup experiment (10-6M): 111
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 8.2
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: 4.27
Experiment series 3
Protein A protein: OSBPL6
Protein A fragment: 285-294
Protein A site: 14-3-3_pSer290
Protein A modification: (SEP)290
Protein A sequence: LQRTQSAPNF
Protein A construct: LQRTQ(pSer)APNF (crude synthetic peptide)
Protein B protein: SFN
Protein B fragment: 1-248
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.81
Holdup BI standard deviation: 0.11
Immobilized partner concentration in holdup experiment (10-6M): 142
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 4.45
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: 4.49
Experiment series 4
Protein A protein: OSBPL6
Protein A fragment: 39-48
Protein A site: 14-3-3_pSer44
Protein A modification: (SEP)44
Protein A sequence: LERTASSSTE
Protein A construct: LERTA(pSer)SSTE (crude synthetic peptide)
Protein B protein: SFN
Protein B fragment: 1-248
Protein B site: 14-3-3
Protein B modification: 100_uM_FCA
Protein B construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: -0.25
Holdup BI standard deviation: 0.57
Immobilized partner concentration in holdup experiment (10-6M): 111
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.51
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 5
Protein A protein: OSBPL6
Protein A fragment: 39-48
Protein A site: 14-3-3_pSer44
Protein A modification: (SEP)44
Protein A sequence: LERTASSSTE
Protein A construct: LERTA(pSer)SSTE (crude synthetic peptide)
Protein B protein: SFN
Protein B fragment: 1-248
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: -0.08
Holdup BI standard deviation: 0.53
Immobilized partner concentration in holdup experiment (10-6M): 142
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 6
Protein A protein: OSBPL6
Protein A fragment: 39-48
Protein A site: 14-3-3_pSer44
Protein A modification: (SEP)44
Protein A sequence: LERTASSSTE
Protein A construct: LERTA(pSer)SSTE (crude synthetic peptide)
Protein B protein: SFN
Protein B fragment: 1-248
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: -0.01
Holdup BI standard deviation: 0.24
Immobilized partner concentration in holdup experiment (10-6M): 113
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.04
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.