Details of experiments between P31946 and Q6WCQ1
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: MPRIP
Protein B fragment: 988-997
Protein B site: 14-3-3_pSer993
Protein B modification: (SEP)993
Protein B sequence: IMKSKSNPDF
Protein B construct: IMKSK(pSer)NPDF (crude synthetic peptide)
Average holdup BI: 1
Holdup BI standard deviation: 0
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): 10.3
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
Measured pKd: 6.31
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: MPRIP
Protein B fragment: 373-382
Protein B site: 14-3-3_pThr378
Protein B modification: (TPO)378
Protein B sequence: LDRRSTEPSV
Protein B construct: LDRRS(pThr)EPSV (crude synthetic peptide)
Average holdup BI: 0.6
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): 8.27
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: 4.17
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: MPRIP
Protein B fragment: 1011-1020
Protein B site: 14-3-3_pSer1016
Protein B modification: (SEP)1016
Protein B sequence: NIRSKSVIEQ
Protein B construct: NIRSK(pSer)VIEQ (crude synthetic peptide)
Average holdup BI: 0.51
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): 7.83
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: 4.01
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: MPRIP
Protein B fragment: 537-546
Protein B site: 14-3-3_pThr542
Protein B modification: (TPO)542
Protein B sequence: EGRSKTFDWA
Protein B construct: EGRSK(pThr)FDWA (crude synthetic peptide)
Average holdup BI: 0.64
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): 6.43
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: 4.24
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: MPRIP
Protein B fragment: 537-546
Protein B site: 14-3-3_pThr542
Protein B modification: (TPO)542
Protein B sequence: EGRSKTFDWA
Protein B construct: EGRSK(pThr)FDWA (crude synthetic peptide)
Average holdup BI: 0.71
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): 6.41
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 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: MPRIP
Protein B fragment: 1011-1020
Protein B site: 14-3-3_pSer1016
Protein B modification: (SEP)1016
Protein B sequence: NIRSKSVIEQ
Protein B construct: NIRSK(pSer)VIEQ (crude synthetic peptide)
Average holdup BI: 0.51
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): 5.72
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: 3.89
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: MPRIP
Protein B fragment: 373-382
Protein B site: 14-3-3_pThr378
Protein B modification: (TPO)378
Protein B sequence: LDRRSTEPSV
Protein B construct: LDRRS(pThr)EPSV (crude synthetic peptide)
Average holdup BI: 0.6
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): 5.65
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: 4.06
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: MPRIP
Protein B fragment: 988-997
Protein B site: 14-3-3_pSer993
Protein B modification: (SEP)993
Protein B sequence: IMKSKSNPDF
Protein B construct: IMKSK(pSer)NPDF (crude synthetic peptide)
Average holdup BI: 1
Holdup BI standard deviation: 0
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): 3.95
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: 6.36
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: MPRIP
Protein B fragment: 264-273
Protein B site: 14-3-3_pSer269
Protein B modification: (SEP)269
Protein B sequence: ESGYFSLEKT
Protein B construct: ESGYF(pSer)LEKT (crude synthetic peptide)
Average holdup BI: 0.14
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): 3.25
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 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: MPRIP
Protein B fragment: 289-298
Protein B site: 14-3-3_pSer294
Protein B modification: (SEP)294
Protein B sequence: SPPSPSTPNH
Protein B construct: SPPSP(pSer)TPNH (crude synthetic peptide)
Average holdup BI: 0.13
Holdup BI standard deviation: 0.05
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): 3.04
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
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: MPRIP
Protein B fragment: 296-305
Protein B site: 14-3-3_pSer301
Protein B modification: (SEP)301
Protein B sequence: PNHRRSQVIE
Protein B construct: PNHRR(pSer)QVIE (crude synthetic peptide)
Average holdup BI: 0.87
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): 2.06
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: 4.71
Experiment series 12
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: MPRIP
Protein B fragment: 289-298
Protein B site: 14-3-3_pSer294
Protein B modification: (SEP)294
Protein B sequence: SPPSPSTPNH
Protein B construct: SPPSP(pSer)TPNH (crude synthetic peptide)
Average holdup BI: 0.14
Holdup BI standard deviation: 0.08
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): 1.82
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 13
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: MPRIP
Protein B fragment: 264-273
Protein B site: 14-3-3_pSer269
Protein B modification: (SEP)269
Protein B sequence: ESGYFSLEKT
Protein B construct: ESGYF(pSer)LEKT (crude synthetic peptide)
Average holdup BI: 0.18
Holdup BI standard deviation: 0.12
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): 1.61
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 14
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: MPRIP
Protein B fragment: 296-305
Protein B site: 14-3-3_pSer301
Protein B modification: (SEP)301
Protein B sequence: PNHRRSQVIE
Protein B construct: PNHRR(pSer)QVIE (crude synthetic peptide)
Average holdup BI: -0.21
Holdup BI standard deviation: 0.45
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): 0.28
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
The affinity was below the detection threshold of the assay.
Experiment series 15
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: MPRIP
Protein B fragment: 1011-1020
Protein B site: 14-3-3_pSer1016
Protein B modification: (SEP)1016
Protein B sequence: NIRSKSLKEG
Protein B construct: NIRSK(pSer)LKEG (crude synthetic peptide)
Average holdup BI: 0.11
Holdup BI standard deviation: 0.74
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): 0.08
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.