Details of experiments between P63104 and Q8IZ21
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: PHACTR4
Protein B fragment: 459-468
Protein B site: 14-3-3_pSer464
Protein B modification: (SEP)464
Protein B sequence: LGRTRSLPIT
Protein B construct: LGRTR(pSer)LPIT (crude synthetic peptide)
Average holdup BI: 0.97
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): 6.88
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: 5.38
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: PHACTR4
Protein B fragment: 142-151
Protein B site: 14-3-3_pSer147
Protein B modification: (SEP)147
Protein B sequence: KKRLGSTGSQ
Protein B construct: KKRLG(pSer)TGSQ (crude synthetic peptide)
Average holdup BI: -0.22
Holdup BI standard deviation: 0.31
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): 1
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 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: PHACTR4
Protein B fragment: 438-447
Protein B site: 14-3-3_pSer443
Protein B modification: (SEP)443
Protein B sequence: SHRAHSLLFE
Protein B construct: SHRAH(pSer)LLFE (crude synthetic peptide)
Average holdup BI: 0.13
Holdup BI standard deviation: 0.13
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): 0.86
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 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: PHACTR4
Protein B fragment: 623-632
Protein B site: 14-3-3_pSer628
Protein B modification: (SEP)628
Protein B sequence: LTRKLSQRPT
Protein B construct: LTRKL(pSer)QRPT (crude synthetic peptide)
Average holdup BI: 0.15
Holdup BI standard deviation: 0.19
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): 0.77
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 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: PHACTR4
Protein B fragment: 65-74
Protein B site: 14-3-3_pSer70
Protein B modification: (SEP)70
Protein B sequence: LERKISMRKP
Protein B construct: LERKI(pSer)MRKP (crude synthetic peptide)
Average holdup BI: 0.01
Holdup BI standard deviation: 0.09
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.
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: PHACTR4
Protein B fragment: 585-594
Protein B site: 14-3-3_pSer590
Protein B modification: (SEP)590
Protein B sequence: LIRRLSQRPT
Protein B construct: LIRRL(pSer)QRPT (crude synthetic peptide)
Average holdup BI: -0.01
Holdup BI standard deviation: 0.08
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): 0.09
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
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: PHACTR4
Protein B fragment: 265-274
Protein B site: 14-3-3_pSer270
Protein B modification: (SEP)270
Protein B sequence: PAHRNSNPVI
Protein B construct: PAHRN(pSer)NPVI (crude synthetic peptide)
Average holdup BI: 0.01
Holdup BI standard deviation: 0.08
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): 0.09
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
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: PHACTR4
Protein B fragment: 552-561
Protein B site: 14-3-3_pSer557
Protein B modification: (SEP)557
Protein B sequence: LNHRPSEPEL
Protein B construct: LNHRP(pSer)EPEL (crude synthetic peptide)
Average holdup BI: 0.01
Holdup BI standard deviation: 0.08
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): 0.06
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 9
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: PHACTR4
Protein B fragment: 126-135
Protein B site: 14-3-3_pSer131
Protein B modification: (SEP)131
Protein B sequence: PVRLASLRKA
Protein B construct: PVRLA(pSer)LRKA (crude synthetic peptide)
Average holdup BI: 0
Holdup BI standard deviation: 0.11
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): 0.01
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.