Details of experiments between P62258 and Q00537
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: CDK17
Protein B fragment: 200-209
Protein B site: 14-3-3_pThr205
Protein B modification: (TPO)205
Protein B sequence: EGTYATVYKG
Protein B construct: EGTYA(pThr)VYKG (crude synthetic peptide)
Average holdup BI: 0.53
Holdup BI standard deviation: 0.05
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): 6.11
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
Measured pKd: 3.9
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: CDK17
Protein B fragment: 200-209
Protein B site: 14-3-3_pThr205
Protein B modification: (TPO)205
Protein B sequence: EGTYATVYKG
Protein B construct: EGTYA(pThr)VYKG (crude synthetic peptide)
Average holdup BI: 0.47
Holdup BI standard deviation: 0.04
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): 5.99
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: 3.89
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: CDK17
Protein B fragment: 175-184
Protein B site: 14-3-3_pSer180
Protein B modification: (SEP)180
Protein B sequence: RSRRASLSEI
Protein B construct: RSRRA(pSer)LSEI (crude synthetic peptide)
Average holdup BI: 0.53
Holdup BI standard deviation: 0.09
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): 5
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: 3.89
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: CDK17
Protein B fragment: 515-524
Protein B site: 14-3-3_pSer520
Protein B modification: (SEP)520
Protein B sequence: KNRRQSMLF-
Protein B construct: GKNRRQ(pSer)MLF (crude synthetic peptide)
Average holdup BI: 0.7
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): 4.66
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: 4.22
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: CDK17
Protein B fragment: 141-150
Protein B site: 14-3-3_pSer146
Protein B modification: (SEP)146
Protein B sequence: LNKRLSLPAD
Protein B construct: LNKRL(pSer)LPAD (crude synthetic peptide)
Average holdup BI: 0.29
Holdup BI standard deviation: 0.06
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): 4.26
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: 3.46
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: CDK17
Protein B fragment: 70-79
Protein B site: 14-3-3_pSer75
Protein B modification: (SEP)75
Protein B sequence: LRRPHSVIGG
Protein B construct: LRRPH(pSer)VIGG (crude synthetic peptide)
Average holdup BI: 0.42
Holdup BI standard deviation: 0.07
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): 4.19
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: 3.79
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: CDK17
Protein B fragment: 197-206
Protein B site: 14-3-3_pThr202
Protein B modification: (TPO)202
Protein B sequence: KLGEGTYATV
Protein B construct: KLGEG(pThr)YATV (crude synthetic peptide)
Average holdup BI: 0.16
Holdup BI standard deviation: 0.04
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): 4.15
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: 3.13
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: CDK17
Protein B fragment: 141-150
Protein B site: 14-3-3_pSer146
Protein B modification: (SEP)146
Protein B sequence: LNKRLSLPAD
Protein B construct: LNKRL(pSer)LPAD (crude synthetic peptide)
Average holdup BI: 0.21
Holdup BI standard deviation: 0.04
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): 3.87
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: CDK17
Protein B fragment: 197-206
Protein B site: 14-3-3_pThr202
Protein B modification: (TPO)202
Protein B sequence: KLGEGTYATV
Protein B construct: KLGEG(pThr)YATV (crude synthetic peptide)
Average holdup BI: 0.12
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): 3.86
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: CDK17
Protein B fragment: 70-79
Protein B site: 14-3-3_pSer75
Protein B modification: (SEP)75
Protein B sequence: LRRPHSVIGG
Protein B construct: LRRPH(pSer)VIGG (crude synthetic peptide)
Average holdup BI: 0.46
Holdup BI standard deviation: 0.16
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): 2.84
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: 3.78
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: CDK17
Protein B fragment: 515-524
Protein B site: 14-3-3_pSer520
Protein B modification: (SEP)520
Protein B sequence: KNRRQSMLF-
Protein B construct: GKNRRQ(pSer)MLF (crude synthetic peptide)
Average holdup BI: 0.79
Holdup BI standard deviation: 0.14
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): 2.12
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: 4.51
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: CDK17
Protein B fragment: 439-448
Protein B site: 14-3-3_pSer444
Protein B modification: (SEP)444
Protein B sequence: APRLDSEGIE
Protein B construct: APRLD(pSer)EGIE (crude synthetic peptide)
Average holdup BI: -0.06
Holdup BI standard deviation: 0.06
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): 1.17
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 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: CDK17
Protein B fragment: 100-109
Protein B site: 14-3-3_pSer105
Protein B modification: (SEP)105
Protein B sequence: NLKMGSDGES
Protein B construct: NLKMG(pSer)DGES (crude synthetic peptide)
Average holdup BI: 0.12
Holdup BI standard deviation: 0.13
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): 0.89
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: 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: CDK17
Protein B fragment: 439-448
Protein B site: 14-3-3_pSer444
Protein B modification: (SEP)444
Protein B sequence: APRLDSEGIE
Protein B construct: APRLD(pSer)EGIE (crude synthetic peptide)
Average holdup BI: 0.13
Holdup BI standard deviation: 0.25
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): 0.47
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: 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: CDK17
Protein B fragment: 132-141
Protein B site: 14-3-3_pSer137
Protein B modification: (SEP)137
Protein B sequence: IHRRISMEDL
Protein B construct: IHRRI(pSer)MEDL (crude synthetic peptide)
Average holdup BI: 0.17
Holdup BI standard deviation: 0.37
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): 0.24
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 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: CDK17
Protein B fragment: 100-109
Protein B site: 14-3-3_pSer105
Protein B modification: (SEP)105
Protein B sequence: NLKMGSDGES
Protein B construct: NLKMG(pSer)DGES (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): 0.17
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 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: CDK17
Protein B fragment: 87-96
Protein B site: 14-3-3_pSer92
Protein B modification: (SEP)92
Protein B sequence: MPRNGSRLDI
Protein B construct: MPRNG(pSer)RLDI (crude synthetic peptide)
Average holdup BI: 0.03
Holdup BI standard deviation: 0.72
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): 0.02
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 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: CDK17
Protein B fragment: 4-13
Protein B site: 14-3-3_pSer9
Protein B modification: (SEP)9
Protein B sequence: FKRRLSLTLR
Protein B construct: FKRRL(pSer)LTLR (crude synthetic peptide)
Average holdup BI: -0.03
Holdup BI standard deviation: 1.09
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.
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: CDK17
Protein B fragment: 132-141
Protein B site: 14-3-3_pSer137
Protein B modification: (SEP)137
Protein B sequence: IHRRISMEDL
Protein B construct: IHRRI(pSer)MEDL (crude synthetic peptide)
Average holdup BI: 0
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
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.