Details of experiments between P31946 and Q5VZ89
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
Protein B protein: DENND4C
Protein B fragment: 1657-1666
Protein B site: 14-3-3_pSer1662
Protein B modification: (SEP)1662
Protein B sequence: GISTVSLPNS
Protein B construct: GISTV(pSer)LPNS (crude synthetic peptide)
Average holdup BI: 0.79
Holdup BI standard deviation: 0.04
Immobilized partner concentration in holdup experiment (10-6M): 102
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 7.93
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: 4.57
Experiment series 2
Protein A protein: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
Protein B protein: DENND4C
Protein B fragment: 1320-1329
Protein B site: 14-3-3_pSer1325
Protein B modification: (SEP)1325
Protein B sequence: KERSTSLSAL
Protein B construct: KERST(pSer)LSAL (crude synthetic peptide)
Average holdup BI: 0.75
Holdup BI standard deviation: 0.02
Immobilized partner concentration in holdup experiment (10-6M): 102
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 7.42
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: 4.47
Experiment series 3
Protein A protein: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
Protein B protein: DENND4C
Protein B fragment: 1320-1329
Protein B site: 14-3-3_pSer1325
Protein B modification: (SEP)1325
Protein B sequence: KERSTSLSAL
Protein B construct: KERST(pSer)LSAL (crude synthetic peptide)
Average holdup BI: 0.75
Holdup BI standard deviation: 0.02
Immobilized partner concentration in holdup experiment (10-6M): 134
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 7.34
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: 4.36
Experiment series 4
Protein A protein: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
Protein B protein: DENND4C
Protein B fragment: 1657-1666
Protein B site: 14-3-3_pSer1662
Protein B modification: (SEP)1662
Protein B sequence: GISTVSLPNS
Protein B construct: GISTV(pSer)LPNS (crude synthetic peptide)
Average holdup BI: 0.77
Holdup BI standard deviation: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 134
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 6.57
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: 4.39
Experiment series 5
Protein A protein: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
Protein B protein: DENND4C
Protein B fragment: 1273-1282
Protein B site: 14-3-3_pSer1278
Protein B modification: (SEP)1278
Protein B sequence: LERRSSLPLD
Protein B construct: LERRS(pSer)LPLD (crude synthetic peptide)
Average holdup BI: 0.42
Holdup BI standard deviation: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 102
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 6.47
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: 3.85
Experiment series 6
Protein A protein: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
Protein B protein: DENND4C
Protein B fragment: 1635-1644
Protein B site: 14-3-3_pSer1640
Protein B modification: (SEP)1640
Protein B sequence: LTRSHSVGGP
Protein B construct: LTRSH(pSer)VGGP (crude synthetic peptide)
Average holdup BI: 0.64
Holdup BI standard deviation: 0.06
Immobilized partner concentration in holdup experiment (10-6M): 102
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 6.46
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: 4.24
Experiment series 7
Protein A protein: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
Protein B protein: DENND4C
Protein B fragment: 1618-1627
Protein B site: 14-3-3_pSer1623
Protein B modification: (SEP)1623
Protein B sequence: TMKISSVPNS
Protein B construct: TMKIS(pSer)VPNS (crude synthetic peptide)
Average holdup BI: 0.48
Holdup BI standard deviation: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 102
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 6.36
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
Measured pKd: 3.96
Experiment series 8
Protein A protein: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
Protein B protein: DENND4C
Protein B fragment: 1273-1282
Protein B site: 14-3-3_pSer1278
Protein B modification: (SEP)1278
Protein B sequence: LERRSSLPLD
Protein B construct: LERRS(pSer)LPLD (crude synthetic peptide)
Average holdup BI: 0.42
Holdup BI standard deviation: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 134
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 6.26
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
Measured pKd: 3.73
Experiment series 9
Protein A protein: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
Protein B protein: DENND4C
Protein B fragment: 1618-1627
Protein B site: 14-3-3_pSer1623
Protein B modification: (SEP)1623
Protein B sequence: TMKISSVPNS
Protein B construct: TMKIS(pSer)VPNS (crude synthetic peptide)
Average holdup BI: 0.58
Holdup BI standard deviation: 0.04
Immobilized partner concentration in holdup experiment (10-6M): 134
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 4.92
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: 4.01
Experiment series 10
Protein A protein: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
Protein B protein: DENND4C
Protein B fragment: 1635-1644
Protein B site: 14-3-3_pSer1640
Protein B modification: (SEP)1640
Protein B sequence: LTRSHSVGGP
Protein B construct: LTRSH(pSer)VGGP (crude synthetic peptide)
Average holdup BI: 0.58
Holdup BI standard deviation: 0.12
Immobilized partner concentration in holdup experiment (10-6M): 134
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 4.1
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: 4.02
Experiment series 11
Protein A protein: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
Protein B protein: DENND4C
Protein B fragment: 1179-1188
Protein B site: 14-3-3_pSer1184
Protein B modification: (SEP)1184
Protein B sequence: RNKRSSLYGI
Protein B construct: RNKRS(pSer)LYGI (crude synthetic peptide)
Average holdup BI: 0.29
Holdup BI standard deviation: 0.07
Immobilized partner concentration in holdup experiment (10-6M): 102
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 4.07
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
Measured pKd: 3.61
Experiment series 12
Protein A protein: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
Protein B protein: DENND4C
Protein B fragment: 1179-1188
Protein B site: 14-3-3_pSer1184
Protein B modification: (SEP)1184
Protein B sequence: RNKRSSLYGI
Protein B construct: RNKRS(pSer)LYGI (crude synthetic peptide)
Average holdup BI: 0.46
Holdup BI standard deviation: 0.17
Immobilized partner concentration in holdup experiment (10-6M): 134
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 2.74
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: 3.8
Experiment series 13
Protein A protein: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
Protein B protein: DENND4C
Protein B fragment: 1094-1103
Protein B site: 14-3-3_pSer1099
Protein B modification: (SEP)1099
Protein B sequence: IARTHSFENV
Protein B construct: IARTH(pSer)FENV (crude synthetic peptide)
Average holdup BI: 0.19
Holdup BI standard deviation: 0.07
Immobilized partner concentration in holdup experiment (10-6M): 134
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 2.3
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: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
Protein B protein: DENND4C
Protein B fragment: 1094-1103
Protein B site: 14-3-3_pSer1099
Protein B modification: (SEP)1099
Protein B sequence: IARTHSFENV
Protein B construct: IARTH(pSer)FENV (crude synthetic peptide)
Average holdup BI: 0.1
Holdup BI standard deviation: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 102
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 2.08
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 15
Protein A protein: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
Protein B protein: DENND4C
Protein B fragment: 1661-1670
Protein B site: 14-3-3_pSer1666
Protein B modification: (SEP)1666
Protein B sequence: VSLPNSLQEV
Protein B construct: VSLPN(pSer)LQEV (crude synthetic peptide)
Average holdup BI: 0.17
Holdup BI standard deviation: 0.16
Immobilized partner concentration in holdup experiment (10-6M): 102
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 1.14
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: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
Protein B protein: DENND4C
Protein B fragment: 948-957
Protein B site: 14-3-3_pSer953
Protein B modification: (SEP)953
Protein B sequence: LIRLESIDNH
Protein B construct: LIRLE(pSer)IDNH (crude synthetic peptide)
Average holdup BI: -0.07
Holdup BI standard deviation: 0.1
Immobilized partner concentration in holdup experiment (10-6M): 102
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.79
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: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
Protein B protein: DENND4C
Protein B fragment: 960-969
Protein B site: 14-3-3_pSer965
Protein B modification: (SEP)965
Protein B sequence: IVAKHSQPSP
Protein B construct: IVAKH(pSer)QPSP (crude synthetic peptide)
Average holdup BI: -0.1
Holdup BI standard deviation: 0.15
Immobilized partner concentration in holdup experiment (10-6M): 134
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.66
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: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
Protein B protein: DENND4C
Protein B fragment: 984-993
Protein B site: 14-3-3_pSer989
Protein B modification: (SEP)989
Protein B sequence: IVKVPSGIFD
Protein B construct: IVKVP(pSer)GIFD (crude synthetic peptide)
Average holdup BI: -0.16
Holdup BI standard deviation: 0.34
Immobilized partner concentration in holdup experiment (10-6M): 102
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.52
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
The affinity was below the detection threshold of the assay.
Experiment series 19
Protein A protein: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
Protein B protein: DENND4C
Protein B fragment: 984-993
Protein B site: 14-3-3_pSer989
Protein B modification: (SEP)989
Protein B sequence: IVKVPSGIFD
Protein B construct: IVKVP(pSer)GIFD (crude synthetic peptide)
Average holdup BI: -0.12
Holdup BI standard deviation: 0.22
Immobilized partner concentration in holdup experiment (10-6M): 134
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.48
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 20
Protein A protein: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
Protein B protein: DENND4C
Protein B fragment: 1341-1350
Protein B site: 14-3-3_pSer1346
Protein B modification: (SEP)1346
Protein B sequence: VVNSLSGLKL
Protein B construct: VVNSL(pSer)GLKL (crude synthetic peptide)
Average holdup BI: 0.06
Holdup BI standard deviation: 0.18
Immobilized partner concentration in holdup experiment (10-6M): 134
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 21
Protein A protein: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
Protein B protein: DENND4C
Protein B fragment: 1661-1670
Protein B site: 14-3-3_pSer1666
Protein B modification: (SEP)1666
Protein B sequence: VSLPNSLQEV
Protein B construct: VSLPN(pSer)LQEV (crude synthetic peptide)
Average holdup BI: 0.14
Holdup BI standard deviation: 0.42
Immobilized partner concentration in holdup experiment (10-6M): 134
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 22
Protein A protein: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
Protein B protein: DENND4C
Protein B fragment: 960-969
Protein B site: 14-3-3_pSer965
Protein B modification: (SEP)965
Protein B sequence: IVAKHSQPSP
Protein B construct: IVAKH(pSer)QPSP (crude synthetic peptide)
Average holdup BI: 0.03
Holdup BI standard deviation: 0.1
Immobilized partner concentration in holdup experiment (10-6M): 102
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.22
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 23
Protein A protein: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
Protein B protein: DENND4C
Protein B fragment: 948-957
Protein B site: 14-3-3_pSer953
Protein B modification: (SEP)953
Protein B sequence: LIRLESIDNH
Protein B construct: LIRLE(pSer)IDNH (crude synthetic peptide)
Average holdup BI: 0.04
Holdup BI standard deviation: 0.2
Immobilized partner concentration in holdup experiment (10-6M): 134
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: 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 24
Protein A protein: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
Protein B protein: DENND4C
Protein B fragment: 1341-1350
Protein B site: 14-3-3_pSer1346
Protein B modification: (SEP)1346
Protein B sequence: VVNSLSGLKL
Protein B construct: VVNSL(pSer)GLKL (crude synthetic peptide)
Average holdup BI: -0.02
Holdup BI standard deviation: 0.26
Immobilized partner concentration in holdup experiment (10-6M): 102
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 25
Protein A protein: YWHAB
Protein A fragment: 1-246
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3beta
Protein B protein: DENND4C
Protein B fragment: 1320-1329
Protein B site: 14-3-3_pSer1325
Protein B modification: (SEP)1325
Protein B sequence: KERSTSLSAL
Protein B construct: biotin-ttds-KERST(pSer)LSAL (HPLC-purified synthetic peptide)
Measured dissociation constant (10-6M): 68
Standard deviation of dissociation constant (10-6M): 4
Experiment method: competitive Fluorescence Polarization
Details of affinity fitting: Competitive binding equation in ProFit
Date of measurement: 2022.02.08
Number of measurements: 3
Measured pKd: 4.17