Details of experiments between Q8TF72 and P27348
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: SHROOM3
Protein A fragment: 414-423
Protein A site: 14-3-3_pSer419
Protein A modification: (SEP)419
Protein A sequence: RPQANSLGSL
Protein A construct: RPQAN(pSer)LGSL (crude synthetic peptide)
Protein B protein: YWHAQ
Protein B fragment: 1-245
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3tau
Average holdup BI: 0.62
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): 7.75
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: 4.1
Experiment series 2
Protein A protein: SHROOM3
Protein A fragment: 1457-1466
Protein A site: 14-3-3_pSer1462
Protein A modification: (SEP)1462
Protein A sequence: YQKQQSLPSL
Protein A construct: YQKQQ(pSer)LPSL (crude synthetic peptide)
Protein B protein: YWHAQ
Protein B fragment: 1-245
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3tau
Average holdup BI: 0.75
Holdup BI standard deviation: 0.03
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): 6.28
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: 4.36
Experiment series 3
Protein A protein: SHROOM3
Protein A fragment: 1216-1225
Protein A site: 14-3-3_pSer1221
Protein A modification: (SEP)1221
Protein A sequence: AGKSMSAEDL
Protein A construct: AGKSM(pSer)AEDL (crude synthetic peptide)
Protein B protein: YWHAQ
Protein B fragment: 1-245
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3tau
Average holdup BI: 0.26
Holdup BI standard deviation: 0.03
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): 5.77
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
Measured pKd: 3.44
Experiment series 4
Protein A protein: SHROOM3
Protein A fragment: 618-627
Protein A site: 14-3-3_pSer623
Protein A modification: (SEP)623
Protein A sequence: RSSRLSEPWE
Protein A construct: RSSRL(pSer)EPWE (crude synthetic peptide)
Protein B protein: YWHAQ
Protein B fragment: 1-245
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3tau
Average holdup BI: 0.31
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): 5.65
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: 3.53
Experiment series 5
Protein A protein: SHROOM3
Protein A fragment: 1416-1425
Protein A site: 14-3-3_pSer1421
Protein A modification: (SEP)1421
Protein A sequence: TRKRVSLPQW
Protein A construct: TRKRV(pSer)LPQW (crude synthetic peptide)
Protein B protein: YWHAQ
Protein B fragment: 1-245
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3tau
Average holdup BI: 0.61
Holdup BI standard deviation: 0.28
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): 2.13
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: 4.07
Experiment series 6
Protein A protein: SHROOM3
Protein A fragment: 1436-1445
Protein A site: 14-3-3_pSer1441
Protein A modification: (SEP)1441
Protein A sequence: AAREDSLPEE
Protein A construct: AARED(pSer)LPEE (crude synthetic peptide)
Protein B protein: YWHAQ
Protein B fragment: 1-245
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3tau
Average holdup BI: -0.1
Holdup BI standard deviation: 0.13
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): 1.06
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 7
Protein A protein: SHROOM3
Protein A fragment: 107-116
Protein A site: 14-3-3_pThr112
Protein A modification: (TPO)112
Protein A sequence: RRDVCTDPGH
Protein A construct: RRDVC(pThr)DPGH (crude synthetic peptide)
Protein B protein: YWHAQ
Protein B fragment: 1-245
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3tau
Average holdup BI: -0.01
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): 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 8
Protein A protein: SHROOM3
Protein A fragment: 1016-1025
Protein A site: 14-3-3_pSer1021
Protein A modification: (SEP)1021
Protein A sequence: KKRSYSEPEK
Protein A construct: KKRSY(pSer)EPEK (crude synthetic peptide)
Protein B protein: YWHAQ
Protein B fragment: 1-245
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3tau
Average holdup BI: 0.01
Holdup BI standard deviation: 0.07
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): 0.06
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: SHROOM3
Protein A fragment: 949-958
Protein A site: 14-3-3_pSer954
Protein A modification: (SEP)954
Protein A sequence: GAPVASRSWR
Protein A construct: GAPVA(pSer)RSWR (crude synthetic peptide)
Protein B protein: YWHAQ
Protein B fragment: 1-245
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3tau
Average holdup BI: -0.01
Holdup BI standard deviation: 0.1
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): 0.05
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: SHROOM3
Protein A fragment: 716-725
Protein A site: 14-3-3_pSer721
Protein A modification: (SEP)721
Protein A sequence: LDRQVSYPRP
Protein A construct: LDRQV(pSer)YPRP (crude synthetic peptide)
Protein B protein: YWHAQ
Protein B fragment: 1-245
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3tau
Average holdup BI: 0.01
Holdup BI standard deviation: 0.15
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): 0.03
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.