Details of experiments between P63104 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: YWHAZ
Protein A fragment: 1-245
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3zeta
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): 132
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 11.96
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: 4.35
Experiment series 2
Protein A protein: YWHAZ
Protein A fragment: 1-245
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3zeta
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: 0.94
Holdup BI standard deviation: 0
Immobilized partner concentration in holdup experiment (10-6M): 132
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 8.9
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: 5.11
Experiment series 3
Protein A protein: YWHAZ
Protein A fragment: 1-245
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3zeta
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: 0.66
Holdup BI standard deviation: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 132
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 7.62
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: 4.16
Experiment series 4
Protein A protein: YWHAZ
Protein A fragment: 1-245
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3zeta
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: 0.85
Holdup BI standard deviation: 0.04
Immobilized partner concentration in holdup experiment (10-6M): 132
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 5.93
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: 4.62
Experiment series 5
Protein A protein: YWHAZ
Protein A fragment: 1-245
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3zeta
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: -0.2
Holdup BI standard deviation: 0.54
Immobilized partner concentration in holdup experiment (10-6M): 132
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.39
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: YWHAZ
Protein A fragment: 1-245
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3zeta
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: -0.13
Holdup BI standard deviation: 1.01
Immobilized partner concentration in holdup experiment (10-6M): 132
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: 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.