Details of experiments between P62258 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: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon
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: 0.92
Holdup BI standard deviation: 0.01
Immobilized partner concentration in holdup experiment (10-6M): 116
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 7.03
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
Experiment series 2
Protein A protein: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon
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: 0.94
Holdup BI standard deviation: 0.01
Immobilized partner concentration in holdup experiment (10-6M): 143
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 6.87
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: 5.02
Experiment series 3
Protein A protein: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon
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: 0.04
Holdup BI standard deviation: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 116
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 1.61
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 4
Protein A protein: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Protein B protein: PHACTR4
Protein B fragment: 112-121
Protein B site: 14-3-3_pSer117
Protein B modification: (SEP)117
Protein B sequence: GNARSSSPVQ
Protein B construct: GNARS(pSer)SPVQ (crude synthetic peptide)
Average holdup BI: 0.3
Holdup BI standard deviation: 0.19
Immobilized partner concentration in holdup experiment (10-6M): 116
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 1.32
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: 3.56
Experiment series 5
Protein A protein: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon
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: -0.11
Holdup BI standard deviation: 0.11
Immobilized partner concentration in holdup experiment (10-6M): 116
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 1.05
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 6
Protein A protein: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon
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: -0.09
Holdup BI standard deviation: 0.12
Immobilized partner concentration in holdup experiment (10-6M): 143
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.81
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: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon
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: -0.16
Holdup BI standard deviation: 0.23
Immobilized partner concentration in holdup experiment (10-6M): 143
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.7
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: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon
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: -0.14
Holdup BI standard deviation: 0.21
Immobilized partner concentration in holdup experiment (10-6M): 116
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.69
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: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon
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: -0.04
Holdup BI standard deviation: 0.1
Immobilized partner concentration in holdup experiment (10-6M): 116
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.4
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 10
Protein A protein: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon
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: -0.05
Holdup BI standard deviation: 0.14
Immobilized partner concentration in holdup experiment (10-6M): 143
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.33
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: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon
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: -0.23
Holdup BI standard deviation: 0.53
Immobilized partner concentration in holdup experiment (10-6M): 116
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.31
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: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon
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: -0.11
Holdup BI standard deviation: 0.34
Immobilized partner concentration in holdup experiment (10-6M): 116
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 13
Protein A protein: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon
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: -0.06
Holdup BI standard deviation: 0.28
Immobilized partner concentration in holdup experiment (10-6M): 143
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 14
Protein A protein: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon
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: -0.04
Holdup BI standard deviation: 0.18
Immobilized partner concentration in holdup experiment (10-6M): 143
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 15
Protein A protein: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon
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: 0.02
Holdup BI standard deviation: 0.08
Immobilized partner concentration in holdup experiment (10-6M): 143
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.17
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: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon
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: -0.04
Holdup BI standard deviation: 0.19
Immobilized partner concentration in holdup experiment (10-6M): 143
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.16
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 17
Protein A protein: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Protein B protein: PHACTR4
Protein B fragment: 112-121
Protein B site: 14-3-3_pSer117
Protein B modification: (SEP)117
Protein B sequence: GNARSSSPVQ
Protein B construct: GNARS(pSer)SPVQ (crude synthetic peptide)
Average holdup BI: -0.15
Holdup BI standard deviation: 1.08
Immobilized partner concentration in holdup experiment (10-6M): 143
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.12
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 18
Protein A protein: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon
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: 0.02
Holdup BI standard deviation: 0.3
Immobilized partner concentration in holdup experiment (10-6M): 116
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.05
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 19
Protein A protein: YWHAE
Protein A fragment: 1-255
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3epsilon
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: 0.01
Holdup BI standard deviation: 0.35
Immobilized partner concentration in holdup experiment (10-6M): 143
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.02
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.