Details of experiments between P31946 and Q53ET0
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
Protein B protein: CRTC2
Protein B fragment: 166-175
Protein B site: 14-3-3_pSer171
Protein B modification: (SEP)171
Protein B sequence: LNRTSSDSAL
Protein B construct: LNRTS(pSer)DSAL (crude synthetic peptide)
Average holdup BI: 0.43
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): 8.13
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: 3.74
Experiment series 2
Protein A protein: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
Protein B protein: CRTC2
Protein B fragment: 122-131
Protein B site: 14-3-3_pSer127
Protein B modification: (SEP)127
Protein B sequence: TRHIDSSPYS
Protein B construct: TRHID(pSer)SPYS (crude synthetic peptide)
Average holdup BI: 0.4
Holdup BI standard deviation: 0.04
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): 6.11
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: 3.82
Experiment series 3
Protein A protein: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
Protein B protein: CRTC2
Protein B fragment: 65-74
Protein B site: 14-3-3_pSer70
Protein B modification: (SEP)70
Protein B sequence: SHYGGSLPNV
Protein B construct: SHYGG(pSer)LPNV (crude synthetic peptide)
Average holdup BI: 0.79
Holdup BI standard deviation: 0.1
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): 5.68
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: 4.56
Experiment series 4
Protein A protein: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
Protein B protein: CRTC2
Protein B fragment: 122-131
Protein B site: 14-3-3_pSer127
Protein B modification: (SEP)127
Protein B sequence: TRHIDSSPYS
Protein B construct: TRHID(pSer)SPYS (crude synthetic peptide)
Average holdup BI: 0.39
Holdup BI standard deviation: 0.04
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): 4.77
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: 3.68
Experiment series 5
Protein A protein: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
Protein B protein: CRTC2
Protein B fragment: 166-175
Protein B site: 14-3-3_pSer171
Protein B modification: (SEP)171
Protein B sequence: LNRTSSDSAL
Protein B construct: LNRTS(pSer)DSAL (crude synthetic peptide)
Average holdup BI: 0.39
Holdup BI standard deviation: 0.07
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): 4.03
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
Measured pKd: 3.8
Experiment series 6
Protein A protein: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
Protein B protein: CRTC2
Protein B fragment: 65-74
Protein B site: 14-3-3_pSer70
Protein B modification: (SEP)70
Protein B sequence: SHYGGSLPNV
Protein B construct: SHYGG(pSer)LPNV (crude synthetic peptide)
Average holdup BI: 0.79
Holdup BI standard deviation: 0.15
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
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: 4.46
Experiment series 7
Protein A protein: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
Protein B protein: CRTC2
Protein B fragment: 487-496
Protein B site: 14-3-3_pSer492
Protein B modification: (SEP)492
Protein B sequence: PYSSPSLVLP
Protein B construct: PYSSP(pSer)LVLP (crude synthetic peptide)
Average holdup BI: 0.17
Holdup BI standard deviation: 0.14
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): 1.25
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: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
Protein B protein: CRTC2
Protein B fragment: 159-168
Protein B site: 14-3-3_pSer164
Protein B modification: (SEP)164
Protein B sequence: LFRLPSALNR
Protein B construct: LFRLP(pSer)ALNR (crude synthetic peptide)
Average holdup BI: -0.12
Holdup BI standard deviation: 0.52
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): 0.21
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: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
Protein B protein: CRTC2
Protein B fragment: 159-168
Protein B site: 14-3-3_pSer164
Protein B modification: (SEP)164
Protein B sequence: LFRLPSALNR
Protein B construct: LFRLP(pSer)ALNR (crude synthetic peptide)
Average holdup BI: 0.16
Holdup BI standard deviation: 0.6
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): 0.2
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: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
Protein B protein: CRTC2
Protein B fragment: 442-451
Protein B site: 14-3-3_pSer447
Protein B modification: (SEP)447
Protein B sequence: ADARRSQQQL
Protein B construct: ADARR(pSer)QQQL (crude synthetic peptide)
Average holdup BI: -0.46
Holdup BI standard deviation: 2.61
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): 0.14
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
The affinity was below the detection threshold of the assay.
Experiment series 11
Protein A protein: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
Protein B protein: CRTC2
Protein B fragment: 442-451
Protein B site: 14-3-3_pSer447
Protein B modification: (SEP)447
Protein B sequence: ADARRSQQQL
Protein B construct: ADARR(pSer)QQQL (crude synthetic peptide)
Average holdup BI: 0.1
Holdup BI standard deviation: 0.59
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): 0.11
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.