Details of experiments between P27348 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: YWHAQ
Protein A fragment: 1-245
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3tau
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: 0.68
Holdup BI standard deviation: 0.02
Immobilized partner concentration in holdup experiment (10-6M): 130
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 7.3
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: 4.21
Experiment series 2
Protein A protein: YWHAQ
Protein A fragment: 1-245
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3tau
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: 0.85
Holdup BI standard deviation: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 130
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 6.01
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
Measured pKd: 4.64
Experiment series 3
Protein A protein: YWHAQ
Protein A fragment: 1-245
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3tau
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: 0.42
Holdup BI standard deviation: 0.06
Immobilized partner concentration in holdup experiment (10-6M): 130
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 5.45
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: 3.75
Experiment series 4
Protein A protein: YWHAQ
Protein A fragment: 1-245
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3tau
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: 0.41
Holdup BI standard deviation: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 130
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 5.16
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
Measured pKd: 3.73
Experiment series 5
Protein A protein: YWHAQ
Protein A fragment: 1-245
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3tau
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: 0.18
Holdup BI standard deviation: 0.04
Immobilized partner concentration in holdup experiment (10-6M): 130
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 4.52
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: 3.23
Experiment series 6
Protein A protein: YWHAQ
Protein A fragment: 1-245
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3tau
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: 0.52
Holdup BI standard deviation: 0.19
Immobilized partner concentration in holdup experiment (10-6M): 130
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 2.66
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: 3.92
Experiment series 7
Protein A protein: YWHAQ
Protein A fragment: 1-245
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3tau
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): 130
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 1.3
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: YWHAQ
Protein A fragment: 1-245
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3tau
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: 0.06
Holdup BI standard deviation: 0.09
Immobilized partner concentration in holdup experiment (10-6M): 130
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.58
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: YWHAQ
Protein A fragment: 1-245
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3tau
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: -0.07
Holdup BI standard deviation: 0.19
Immobilized partner concentration in holdup experiment (10-6M): 130
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.38
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
The affinity was below the detection threshold of the assay.
Experiment series 10
Protein A protein: YWHAQ
Protein A fragment: 1-245
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3tau
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: 0.14
Holdup BI standard deviation: 0.34
Immobilized partner concentration in holdup experiment (10-6M): 130
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.36
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.