Details of experiments between Q13009 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: TIAM1
Protein A fragment: 55-64
Protein A site: 14-3-3_pSer60
Protein A modification: (SEP)60
Protein A sequence: STRSSSTPSI
Protein A construct: STRSS(pSer)TPSI (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: 0.58
Holdup BI standard deviation: 0.01
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): 9.51
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
Measured pKd: 4.09
Experiment series 2
Protein A protein: TIAM1
Protein A fragment: 55-64
Protein A site: 14-3-3_pSer60
Protein A modification: (SEP)60
Protein A sequence: STRSSSTPSI
Protein A construct: STRSS(pSer)TPSI (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: 0.65
Holdup BI standard deviation: 0.03
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): 9.28
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
Measured pKd: 4.12
Experiment series 3
Protein A protein: TIAM1
Protein A fragment: 1432-1441
Protein A site: 14-3-3_pSer1437
Protein A modification: (SEP)1437
Protein A sequence: MSRAVSAPSK
Protein A construct: MSRAV(pSer)APSK (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: 0.95
Holdup BI standard deviation: 0.01
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): 9.09
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: 5.12
Experiment series 4
Protein A protein: TIAM1
Protein A fragment: 55-64
Protein A site: 14-3-3_pSer60
Protein A modification: (SEP)60
Protein A sequence: STRSSSTPSI
Protein A construct: STRSS(pSer)TPSI (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: 0.01
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): 9.08
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: 4.25
Experiment series 5
Protein A protein: TIAM1
Protein A fragment: 1402-1411
Protein A site: 14-3-3_pSer1407
Protein A modification: (SEP)1407
Protein A sequence: LLKTESLPSS
Protein A construct: LLKTE(pSer)LPSS (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: 0.4
Holdup BI standard deviation: 0.02
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): 6.91
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: 3.77
Experiment series 6
Protein A protein: TIAM1
Protein A fragment: 1402-1411
Protein A site: 14-3-3_pSer1407
Protein A modification: (SEP)1407
Protein A sequence: LLKTESLPSS
Protein A construct: LLKTE(pSer)LPSS (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: 0.47
Holdup BI standard deviation: 0.05
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): 5.83
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: 3.79
Experiment series 7
Protein A protein: TIAM1
Protein A fragment: 1432-1441
Protein A site: 14-3-3_pSer1437
Protein A modification: (SEP)1437
Protein A sequence: MSRAVSAPSK
Protein A construct: MSRAV(pSer)APSK (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: 0.94
Holdup BI standard deviation: 0.01
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): 5.43
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: 5.16
Experiment series 8
Protein A protein: TIAM1
Protein A fragment: 314-323
Protein A site: 14-3-3_pThr319
Protein A modification: (TPO)319
Protein A sequence: GRRAKTTQDV
Protein A construct: GRRAK(pThr)TQDV (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: 0.71
Holdup BI standard deviation: 0.07
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): 5.08
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
Measured pKd: 4.24
Experiment series 9
Protein A protein: TIAM1
Protein A fragment: 1402-1411
Protein A site: 14-3-3_pSer1407
Protein A modification: (SEP)1407
Protein A sequence: LLKTESLPSS
Protein A construct: LLKTE(pSer)LPSS (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: 0.38
Holdup BI standard deviation: 0.04
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): 4.94
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 10
Protein A protein: TIAM1
Protein A fragment: 1432-1441
Protein A site: 14-3-3_pSer1437
Protein A modification: (SEP)1437
Protein A sequence: MSRAVSAPSK
Protein A construct: MSRAV(pSer)APSK (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: 0.89
Holdup BI standard deviation: 0.04
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): 3.16
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: 4.84
Experiment series 11
Protein A protein: TIAM1
Protein A fragment: 329-338
Protein A site: 14-3-3_pSer334
Protein A modification: (SEP)334
Protein A sequence: SEFADSGIEG
Protein A construct: SEFAD(pSer)GIEG (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: 0.19
Holdup BI standard deviation: 0.36
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): 0.48
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
The affinity was below the detection threshold of the assay.
Experiment series 12
Protein A protein: TIAM1
Protein A fragment: 690-699
Protein A site: 14-3-3_pSer695
Protein A modification: (SEP)695
Protein A sequence: RSRFSSLWGL
Protein A construct: RSRFS(pSer)LWGL (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: 0.21
Holdup BI standard deviation: 0.63
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): 0.26
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 13
Protein A protein: TIAM1
Protein A fragment: 314-323
Protein A site: 14-3-3_pThr319
Protein A modification: (TPO)319
Protein A sequence: GRRAKTTQDV
Protein A construct: GRRAK(pThr)TQDV (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: -0.21
Holdup BI standard deviation: 0.75
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): 0.23
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 14
Protein A protein: TIAM1
Protein A fragment: 167-176
Protein A site: 14-3-3_pSer172
Protein A modification: (SEP)172
Protein A sequence: KKRSKSADIW
Protein A construct: KKRSK(pSer)ADIW (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: 0.12
Holdup BI standard deviation: 0.46
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): 0.21
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 15
Protein A protein: TIAM1
Protein A fragment: 329-338
Protein A site: 14-3-3_pSer334
Protein A modification: (SEP)334
Protein A sequence: SEFADSGIEG
Protein A construct: SEFAD(pSer)GIEG (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: -0.1
Holdup BI standard deviation: 0.5
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): 0.18
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 16
Protein A protein: TIAM1
Protein A fragment: 167-176
Protein A site: 14-3-3_pSer172
Protein A modification: (SEP)172
Protein A sequence: KKRSKSADIW
Protein A construct: KKRSK(pSer)ADIW (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: 0.85
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): 0.01
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.