Details of experiments between P63104 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: 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: 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: 0.99
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): 8.38
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: 6.06
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: 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: 0.63
Holdup BI standard deviation: 0.02
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): 8.28
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: 4.11
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: 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: 0.49
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): 7.13
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
Measured pKd: 3.86
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: 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: 0.63
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): 6.26
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
Measured pKd: 4.11
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: 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: 0.15
Holdup BI standard deviation: 0.06
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): 2.82
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: 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: 0.07
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): 2.27
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
The affinity was below the detection threshold of the assay.
Experiment series 7
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: 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: -0.26
Holdup BI standard deviation: 0.72
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): 0.2
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 8
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: 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: -0.04
Holdup BI standard deviation: 0.69
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): 0.04
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.