Details of experiments between P31946 and Q13009
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: TIAM1
Protein B fragment: 55-64
Protein B site: 14-3-3_pSer60
Protein B modification: (SEP)60
Protein B sequence: STRSSSTPSI
Protein B construct: STRSS(pSer)TPSI (crude synthetic peptide)
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): 13.54
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 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: TIAM1
Protein B fragment: 55-64
Protein B site: 14-3-3_pSer60
Protein B modification: (SEP)60
Protein B sequence: STRSSSTPSI
Protein B construct: STRSS(pSer)TPSI (crude synthetic peptide)
Average holdup BI: 0.75
Holdup BI standard deviation: 0.01
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): 10.22
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: 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: TIAM1
Protein B fragment: 1432-1441
Protein B site: 14-3-3_pSer1437
Protein B modification: (SEP)1437
Protein B sequence: MSRAVSAPSK
Protein B construct: MSRAV(pSer)APSK (crude synthetic peptide)
Average holdup BI: 0.95
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): 9.24
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: 5.29
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: TIAM1
Protein B fragment: 1402-1411
Protein B site: 14-3-3_pSer1407
Protein B modification: (SEP)1407
Protein B sequence: LLKTESLPSS
Protein B construct: LLKTE(pSer)LPSS (crude synthetic peptide)
Average holdup BI: 0.72
Holdup BI standard deviation: 0.01
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): 8.93
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: 4.28
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: TIAM1
Protein B fragment: 1402-1411
Protein B site: 14-3-3_pSer1407
Protein B modification: (SEP)1407
Protein B sequence: LLKTESLPSS
Protein B construct: LLKTE(pSer)LPSS (crude synthetic peptide)
Average holdup BI: 0.73
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): 7.67
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: 4.42
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: TIAM1
Protein B fragment: 314-323
Protein B site: 14-3-3_pThr319
Protein B modification: (TPO)319
Protein B sequence: GRRAKTTQDV
Protein B construct: GRRAK(pThr)TQDV (crude synthetic peptide)
Average holdup BI: 0.8
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): 5.64
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 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: TIAM1
Protein B fragment: 1432-1441
Protein B site: 14-3-3_pSer1437
Protein B modification: (SEP)1437
Protein B sequence: MSRAVSAPSK
Protein B construct: MSRAV(pSer)APSK (crude synthetic peptide)
Average holdup BI: 0.96
Holdup BI standard deviation: 0.01
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): 5.48
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: 5.27
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: TIAM1
Protein B fragment: 329-338
Protein B site: 14-3-3_pSer334
Protein B modification: (SEP)334
Protein B sequence: SEFADSGIEG
Protein B construct: SEFAD(pSer)GIEG (crude synthetic peptide)
Average holdup BI: 0.17
Holdup BI standard deviation: 0.52
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): 0.26
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: 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: TIAM1
Protein B fragment: 329-338
Protein B site: 14-3-3_pSer334
Protein B modification: (SEP)334
Protein B sequence: SEFADSGIEG
Protein B construct: SEFAD(pSer)GIEG (crude synthetic peptide)
Average holdup BI: -0.15
Holdup BI standard deviation: 0.59
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): 0.24
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: 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: TIAM1
Protein B fragment: 167-176
Protein B site: 14-3-3_pSer172
Protein B modification: (SEP)172
Protein B sequence: KKRSKSADIW
Protein B construct: KKRSK(pSer)ADIW (crude synthetic peptide)
Average holdup BI: -0.19
Holdup BI standard deviation: 1.07
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): 0.13
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: TIAM1
Protein B fragment: 314-323
Protein B site: 14-3-3_pThr319
Protein B modification: (TPO)319
Protein B sequence: GRRAKTTQDV
Protein B construct: GRRAK(pThr)TQDV (crude synthetic peptide)
Average holdup BI: 0.16
Holdup BI standard deviation: 0.96
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): 0.1
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 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: TIAM1
Protein B fragment: 167-176
Protein B site: 14-3-3_pSer172
Protein B modification: (SEP)172
Protein B sequence: KKRSKSADIW
Protein B construct: KKRSK(pSer)ADIW (crude synthetic peptide)
Average holdup BI: 0.01
Holdup BI standard deviation: 0.69
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): 0.01
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.