Details of experiments between P62258 and O94915
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: FRYL
Protein B fragment: 1909-1918
Protein B site: 14-3-3_pSer1914
Protein B modification: (SEP)1914
Protein B sequence: AANRKSTGQL
Protein B construct: AANRK(pSer)TGQL (crude synthetic peptide)
Measured dissociation constant (10-6M): <100
Immobilized partner concentration in holdup experiment (10-6M): 116
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: 1
Measured pKd: 4
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: FRYL
Protein B fragment: 1973-1982
Protein B site: 14-3-3_pSer1978
Protein B modification: (SEP)1978
Protein B sequence: LARTRSLSSL
Protein B construct: LARTR(pSer)LSSL (crude synthetic peptide)
Average holdup BI: 0.82
Holdup BI standard deviation: 0.02
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): 9.12
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
Measured pKd: 4.51
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: FRYL
Protein B fragment: 1973-1982
Protein B site: 14-3-3_pSer1978
Protein B modification: (SEP)1978
Protein B sequence: LARTRSLSSL
Protein B construct: LARTR(pSer)LSSL (crude synthetic peptide)
Average holdup BI: 0.79
Holdup BI standard deviation: 0.02
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): 5.52
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: 4.52
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: FRYL
Protein B fragment: 1936-1945
Protein B site: 14-3-3_pSer1941
Protein B modification: (SEP)1941
Protein B sequence: NARSNSLRLS
Protein B construct: NARSN(pSer)LRLS (crude synthetic peptide)
Average holdup BI: 0.24
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): 4.77
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 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: FRYL
Protein B fragment: 1954-1963
Protein B site: 14-3-3_pThr1959
Protein B modification: (TPO)1959
Protein B sequence: RRRSNTLDIM
Protein B construct: RRRSN(pThr)LDIM (crude synthetic peptide)
Average holdup BI: 0.76
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): 4.63
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: 4.34
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: FRYL
Protein B fragment: 1936-1945
Protein B site: 14-3-3_pSer1941
Protein B modification: (SEP)1941
Protein B sequence: NARSNSLRLS
Protein B construct: NARSN(pSer)LRLS (crude synthetic peptide)
Average holdup BI: 0.22
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): 3.82
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: 3.3
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: FRYL
Protein B fragment: 1967-1976
Protein B site: 14-3-3_pSer1972
Protein B modification: (SEP)1972
Protein B sequence: INHSSSLART
Protein B construct: INHSS(pSer)LART (crude synthetic peptide)
Average holdup BI: 0.09
Holdup BI standard deviation: 0.05
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): 1.78
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 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: FRYL
Protein B fragment: 1940-1949
Protein B site: 14-3-3_pSer1945
Protein B modification: (SEP)1945
Protein B sequence: NSLRLSLIGD
Protein B construct: NSLRL(pSer)LIGD (crude synthetic peptide)
Average holdup BI: -0.22
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): 0.89
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 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: FRYL
Protein B fragment: 1930-1939
Protein B site: 14-3-3_pSer1935
Protein B modification: (SEP)1935
Protein B sequence: YLGYNSNARS
Protein B construct: YLGYN(pSer)NARS (crude synthetic peptide)
Average holdup BI: 0.05
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): 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 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: FRYL
Protein B fragment: 477-486
Protein B site: 14-3-3_pThr482
Protein B modification: (TPO)482
Protein B sequence: LPSGNTLRVK
Protein B construct: LPSGN(pThr)LRVK (crude synthetic peptide)
Average holdup BI: 0.02
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): 0.68
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 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: FRYL
Protein B fragment: 98-107
Protein B site: 14-3-3_pSer103
Protein B modification: (SEP)103
Protein B sequence: RSSTKSKGDE
Protein B construct: RSSTK(pSer)KGDE (crude synthetic peptide)
Average holdup BI: -0.25
Holdup BI standard deviation: 0.5
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): 0.58
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: FRYL
Protein B fragment: 1930-1939
Protein B site: 14-3-3_pSer1935
Protein B modification: (SEP)1935
Protein B sequence: YLGYNSNARS
Protein B construct: YLGYN(pSer)NARS (crude synthetic peptide)
Average holdup BI: -0.02
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): 0.53
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: FRYL
Protein B fragment: 98-107
Protein B site: 14-3-3_pSer103
Protein B modification: (SEP)103
Protein B sequence: RSSTKSKGDE
Protein B construct: RSSTK(pSer)KGDE (crude synthetic peptide)
Average holdup BI: -0.16
Holdup BI standard deviation: 0.51
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): 0.27
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: FRYL
Protein B fragment: 1954-1963
Protein B site: 14-3-3_pThr1959
Protein B modification: (TPO)1959
Protein B sequence: RRRSNTLDIM
Protein B construct: RRRSN(pThr)LDIM (crude synthetic peptide)
Average holdup BI: -0.2
Holdup BI standard deviation: 1.29
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): 0.13
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: FRYL
Protein B fragment: 1909-1918
Protein B site: 14-3-3_pSer1914
Protein B modification: (SEP)1914
Protein B sequence: AANRKSTGQL
Protein B construct: AANRK(pSer)TGQL (crude synthetic peptide)
Average holdup BI: -0.15
Holdup BI standard deviation: 0.85
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): 0.11
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 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: FRYL
Protein B fragment: 477-486
Protein B site: 14-3-3_pThr482
Protein B modification: (TPO)482
Protein B sequence: LPSGNTLRVK
Protein B construct: LPSGN(pThr)LRVK (crude synthetic peptide)
Average holdup BI: -0.01
Holdup BI standard deviation: 0.1
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): 0.08
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: FRYL
Protein B fragment: 1967-1976
Protein B site: 14-3-3_pSer1972
Protein B modification: (SEP)1972
Protein B sequence: INHSSSLART
Protein B construct: INHSS(pSer)LART (crude synthetic peptide)
Average holdup BI: 0.01
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): 0.07
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: FRYL
Protein B fragment: 2801-2810
Protein B site: 14-3-3_pThr2806
Protein B modification: (TPO)2806
Protein B sequence: DDCKRTFGAK
Protein B construct: DDCKR(pThr)FGAK (crude synthetic peptide)
Average holdup BI: 0.12
Holdup BI standard deviation: 1.59
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.