Details of experiments between P31946 and Q6R327
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: RICTOR
Protein B fragment: 1214-1223
Protein B site: 14-3-3_pSer1219
Protein B modification: (SEP)1219
Protein B sequence: KIRSQSFNTD
Protein B construct: KIRSQ(pSer)FNTD (crude synthetic peptide)
Average holdup BI: 0.55
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): 8.39
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: 4.08
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: RICTOR
Protein B fragment: 1279-1288
Protein B site: 14-3-3_pSer1284
Protein B modification: (SEP)1284
Protein B sequence: LSKSNSVSLV
Protein B construct: LSKSN(pSer)VSLV (crude synthetic peptide)
Average holdup BI: 0.44
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): 7.66
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
Measured pKd: 3.76
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: RICTOR
Protein B fragment: 1214-1223
Protein B site: 14-3-3_pSer1219
Protein B modification: (SEP)1219
Protein B sequence: KIRSQSFNTD
Protein B construct: KIRSQ(pSer)FNTD (crude synthetic peptide)
Average holdup BI: 0.52
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): 7.09
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: 3.91
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: RICTOR
Protein B fragment: 1279-1288
Protein B site: 14-3-3_pSer1284
Protein B modification: (SEP)1284
Protein B sequence: LSKSNSVSLV
Protein B construct: LSKSN(pSer)VSLV (crude synthetic peptide)
Average holdup BI: 0.42
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): 6.5
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: 3.85
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: RICTOR
Protein B fragment: 1297-1306
Protein B site: 14-3-3_pSer1302
Protein B modification: (SEP)1302
Protein B sequence: PRRAQSLKAP
Protein B construct: PRRAQ(pSer)LKAP (crude synthetic peptide)
Average holdup BI: 0.78
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): 5.79
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: 4.43
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: RICTOR
Protein B fragment: 1380-1389
Protein B site: 14-3-3_pSer1385
Protein B modification: (SEP)1385
Protein B sequence: FMKALSYASL
Protein B construct: FMKAL(pSer)YASL (crude synthetic peptide)
Average holdup BI: 0.78
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): 5.7
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: 4.43
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: RICTOR
Protein B fragment: 1297-1306
Protein B site: 14-3-3_pSer1302
Protein B modification: (SEP)1302
Protein B sequence: PRRAQSLKAP
Protein B construct: PRRAQ(pSer)LKAP (crude synthetic peptide)
Average holdup BI: 0.8
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): 5.33
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: 4.59
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: RICTOR
Protein B fragment: 1128-1137
Protein B site: 14-3-3_pThr1133
Protein B modification: (TPO)1133
Protein B sequence: NRRIRTLTEP
Protein B construct: NRRIR(pThr)LTEP (crude synthetic peptide)
Average holdup BI: 0.39
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): 4.9
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: 3.79
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: RICTOR
Protein B fragment: 1383-1392
Protein B site: 14-3-3_pSer1388
Protein B modification: (SEP)1388
Protein B sequence: ALSYASLDKE
Protein B construct: ALSYA(pSer)LDKE (crude synthetic peptide)
Average holdup BI: 0.06
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): 3.49
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: 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: RICTOR
Protein B fragment: 1380-1389
Protein B site: 14-3-3_pSer1385
Protein B modification: (SEP)1385
Protein B sequence: FMKALSYASL
Protein B construct: FMKAL(pSer)YASL (crude synthetic peptide)
Average holdup BI: 0.66
Holdup BI standard deviation: 0.14
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): 2.5
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: 4.29
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: RICTOR
Protein B fragment: 1172-1181
Protein B site: 14-3-3_pSer1177
Protein B modification: (SEP)1177
Protein B sequence: TGSTPSIGEN
Protein B construct: TGSTP(pSer)IGEN (crude synthetic peptide)
Average holdup BI: -0.2
Holdup BI standard deviation: 0.1
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): 2.46
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: 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: RICTOR
Protein B fragment: 1079-1088
Protein B site: 14-3-3_pSer1084
Protein B modification: (SEP)1084
Protein B sequence: IKDKNSFPFF
Protein B construct: IKDKN(pSer)FPFF (crude synthetic peptide)
Average holdup BI: 0.69
Holdup BI standard deviation: 0.21
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): 2.4
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: 4.23
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: RICTOR
Protein B fragment: 1130-1139
Protein B site: 14-3-3_pThr1135
Protein B modification: (TPO)1135
Protein B sequence: RIRTLTEPSV
Protein B construct: RIRTL(pThr)EPSV (crude synthetic peptide)
Average holdup BI: 0.24
Holdup BI standard deviation: 0.1
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): 2.21
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: RICTOR
Protein B fragment: 1079-1088
Protein B site: 14-3-3_pSer1084
Protein B modification: (SEP)1084
Protein B sequence: IKDKNSFPFF
Protein B construct: IKDKN(pSer)FPFF (crude synthetic peptide)
Average holdup BI: 0.59
Holdup BI standard deviation: 0.18
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): 2.2
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: 4.14
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: RICTOR
Protein B fragment: 1128-1137
Protein B site: 14-3-3_pThr1133
Protein B modification: (TPO)1133
Protein B sequence: NRRIRTLTEP
Protein B construct: NRRIR(pThr)LTEP (crude synthetic peptide)
Average holdup BI: 0.38
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): 1.86
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
Measured pKd: 3.65
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: RICTOR
Protein B fragment: 378-387
Protein B site: 14-3-3_pSer383
Protein B modification: (SEP)383
Protein B sequence: PHRARSRPDL
Protein B construct: PHRAR(pSer)RPDL (crude synthetic peptide)
Average holdup BI: 0.26
Holdup BI standard deviation: 0.16
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): 1.74
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: 3.43
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: RICTOR
Protein B fragment: 1130-1139
Protein B site: 14-3-3_pThr1135
Protein B modification: (TPO)1135
Protein B sequence: RIRTLTEPSV
Protein B construct: RIRTL(pThr)EPSV (crude synthetic peptide)
Average holdup BI: 0.31
Holdup BI standard deviation: 0.19
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): 1.64
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: 3.64
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: RICTOR
Protein B fragment: 1172-1181
Protein B site: 14-3-3_pSer1177
Protein B modification: (SEP)1177
Protein B sequence: TGSTPSIGEN
Protein B construct: TGSTP(pSer)IGEN (crude synthetic peptide)
Average holdup BI: -0.16
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): 1.13
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
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: RICTOR
Protein B fragment: 1383-1392
Protein B site: 14-3-3_pSer1388
Protein B modification: (SEP)1388
Protein B sequence: ALSYASLDKE
Protein B construct: ALSYA(pSer)LDKE (crude synthetic peptide)
Average holdup BI: 0.11
Holdup BI standard deviation: 0.15
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): 0.77
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
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: RICTOR
Protein B fragment: 1133-1142
Protein B site: 14-3-3_pSer1138
Protein B modification: (SEP)1138
Protein B sequence: TLTEPSVDFN
Protein B construct: TLTEP(pSer)VDFN (crude synthetic peptide)
Average holdup BI: -0.07
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): 0.33
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: RICTOR
Protein B fragment: 378-387
Protein B site: 14-3-3_pSer383
Protein B modification: (SEP)383
Protein B sequence: PHRARSRPDL
Protein B construct: PHRAR(pSer)RPDL (crude synthetic peptide)
Average holdup BI: 0.05
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): 0.25
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 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: RICTOR
Protein B fragment: 1327-1336
Protein B site: 14-3-3_pThr1332
Protein B modification: (TPO)1332
Protein B sequence: AFGYATLKRL
Protein B construct: AFGYA(pThr)LKRL (crude synthetic peptide)
Average holdup BI: 0.11
Holdup BI standard deviation: 0.36
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): 0.24
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: RICTOR
Protein B fragment: 1133-1142
Protein B site: 14-3-3_pSer1138
Protein B modification: (SEP)1138
Protein B sequence: TLTEPSVDFN
Protein B construct: TLTEP(pSer)VDFN (crude synthetic peptide)
Average holdup BI: -0.03
Holdup BI standard deviation: 0.28
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): 0.09
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 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: RICTOR
Protein B fragment: 1214-1223
Protein B site: 14-3-3_pSer1219
Protein B modification: (SEP)1219
Protein B sequence: KIRSQSFNTD
Protein B construct: biotin-ttds-KIRSQ(pSer)FNTD (HPLC-purified synthetic peptide)
Measured dissociation constant (10-6M): 121
Standard deviation of dissociation constant (10-6M): 38
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: 3.92