Details of experiments between Q12802 and P31946
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: AKAP13
Protein A fragment: 1597-1606
Protein A site: 14-3-3_pSer1602
Protein A modification: (SEP)1602
Protein A sequence: HRRSFSLEGL
Protein A construct: HRRSF(pSer)LEGL (crude synthetic peptide)
Protein B protein: YWHAB
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3beta
Measured dissociation constant (10-6M): <100
Immobilized partner concentration in holdup experiment (10-6M): 102
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: 1
Measured pKd: 4
Experiment series 2
Protein A protein: AKAP13
Protein A fragment: 2462-2471
Protein A site: 14-3-3_pThr2467
Protein A modification: (TPO)2467
Protein A sequence: PRRAETFGGF
Protein A construct: PRRAE(pThr)FGGF (crude synthetic peptide)
Protein B protein: YWHAB
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.97
Holdup BI standard deviation: 0
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): 7.52
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: 5.44
Experiment series 3
Protein A protein: AKAP13
Protein A fragment: 2493-2502
Protein A site: 14-3-3_pSer2498
Protein A modification: (SEP)2498
Protein A sequence: LRRTESDSGL
Protein A construct: LRRTE(pSer)DSGL (crude synthetic peptide)
Protein B protein: YWHAB
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.46
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): 6.14
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: 3.81
Experiment series 4
Protein A protein: AKAP13
Protein A fragment: 1924-1933
Protein A site: 14-3-3_pSer1929
Protein A modification: (SEP)1929
Protein A sequence: KFLSHSTDSL
Protein A construct: KFLSH(pSer)TDSL (crude synthetic peptide)
Protein B protein: YWHAB
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.57
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): 6.05
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
Measured pKd: 4.12
Experiment series 5
Protein A protein: AKAP13
Protein A fragment: 2493-2502
Protein A site: 14-3-3_pSer2498
Protein A modification: (SEP)2498
Protein A sequence: LRRTESDSGL
Protein A construct: LRRTE(pSer)DSGL (crude synthetic peptide)
Protein B protein: YWHAB
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.41
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): 5.82
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: 3.83
Experiment series 6
Protein A protein: AKAP13
Protein A fragment: 2462-2471
Protein A site: 14-3-3_pThr2467
Protein A modification: (TPO)2467
Protein A sequence: PRRAETFGGF
Protein A construct: PRRAE(pThr)FGGF (crude synthetic peptide)
Protein B protein: YWHAB
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.98
Holdup BI standard deviation: 0.01
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): 5.63
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: 5.67
Experiment series 7
Protein A protein: AKAP13
Protein A fragment: 1502-1511
Protein A site: 14-3-3_pSer1507
Protein A modification: (SEP)1507
Protein A sequence: SRDRQSLDGF
Protein A construct: SRDRQ(pSer)LDGF (crude synthetic peptide)
Protein B protein: YWHAB
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.28
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): 5.4
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: 3.46
Experiment series 8
Protein A protein: AKAP13
Protein A fragment: 1924-1933
Protein A site: 14-3-3_pSer1929
Protein A modification: (SEP)1929
Protein A sequence: KFLSHSTDSL
Protein A construct: KFLSH(pSer)TDSL (crude synthetic peptide)
Protein B protein: YWHAB
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.56
Holdup BI standard deviation: 0.06
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): 4.87
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: 3.99
Experiment series 9
Protein A protein: AKAP13
Protein A fragment: 1871-1880
Protein A site: 14-3-3_pSer1876
Protein A modification: (SEP)1876
Protein A sequence: KERPRSAVLL
Protein A construct: KERPR(pSer)AVLL (crude synthetic peptide)
Protein B protein: YWHAB
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.62
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): 4.71
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: 4.2
Experiment series 10
Protein A protein: AKAP13
Protein A fragment: 1745-1754
Protein A site: 14-3-3_pSer1750
Protein A modification: (SEP)1750
Protein A sequence: VSRTFSYIKN
Protein A construct: VSRTF(pSer)YIKN (crude synthetic peptide)
Protein B protein: YWHAB
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.23
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): 4.7
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: 3.47
Experiment series 11
Protein A protein: AKAP13
Protein A fragment: 1502-1511
Protein A site: 14-3-3_pSer1507
Protein A modification: (SEP)1507
Protein A sequence: SRDRQSLDGF
Protein A construct: SRDRQ(pSer)LDGF (crude synthetic peptide)
Protein B protein: YWHAB
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.25
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): 4.28
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: 3.52
Experiment series 12
Protein A protein: AKAP13
Protein A fragment: 2562-2571
Protein A site: 14-3-3_pSer2567
Protein A modification: (SEP)2567
Protein A sequence: LSRPSSLIEQ
Protein A construct: LSRPS(pSer)LIEQ (crude synthetic peptide)
Protein B protein: YWHAB
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.2
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): 4.24
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 13
Protein A protein: AKAP13
Protein A fragment: 1871-1880
Protein A site: 14-3-3_pSer1876
Protein A modification: (SEP)1876
Protein A sequence: KERPRSAVLL
Protein A construct: KERPR(pSer)AVLL (crude synthetic peptide)
Protein B protein: YWHAB
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.71
Holdup BI standard deviation: 0.13
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: 4.25
Experiment series 14
Protein A protein: AKAP13
Protein A fragment: 2406-2415
Protein A site: 14-3-3_pThr2411
Protein A modification: (TPO)2411
Protein A sequence: LFRSNTEEAL
Protein A construct: LFRSN(pThr)EEAL (crude synthetic peptide)
Protein B protein: YWHAB
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.21
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): 3.62
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: AKAP13
Protein A fragment: 2406-2415
Protein A site: 14-3-3_pThr2411
Protein A modification: (TPO)2411
Protein A sequence: LFRSNTEEAL
Protein A construct: LFRSN(pThr)EEAL (crude synthetic peptide)
Protein B protein: YWHAB
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.21
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): 3.39
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
The affinity was below the detection threshold of the assay.
Experiment series 16
Protein A protein: AKAP13
Protein A fragment: 2558-2567
Protein A site: 14-3-3_pSer2563
Protein A modification: (SEP)2563
Protein A sequence: LTRSLSRPSS
Protein A construct: LTRSL(pSer)RPSS (crude synthetic peptide)
Protein B protein: YWHAB
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.3
Holdup BI standard deviation: 0.08
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): 3.33
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
Measured pKd: 3.51
Experiment series 17
Protein A protein: AKAP13
Protein A fragment: 1745-1754
Protein A site: 14-3-3_pSer1750
Protein A modification: (SEP)1750
Protein A sequence: VSRTFSYIKN
Protein A construct: VSRTF(pSer)YIKN (crude synthetic peptide)
Protein B protein: YWHAB
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.34
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): 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
Measured pKd: 3.58
Experiment series 18
Protein A protein: AKAP13
Protein A fragment: 1221-1230
Protein A site: 14-3-3_pSer1226
Protein A modification: (SEP)1226
Protein A sequence: SGRERSTPSL
Protein A construct: SGRER(pSer)TPSL (crude synthetic peptide)
Protein B protein: YWHAB
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.11
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): 2.64
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
The affinity was below the detection threshold of the assay.
Experiment series 19
Protein A protein: AKAP13
Protein A fragment: 2562-2571
Protein A site: 14-3-3_pSer2567
Protein A modification: (SEP)2567
Protein A sequence: LSRPSSLIEQ
Protein A construct: LSRPS(pSer)LIEQ (crude synthetic peptide)
Protein B protein: YWHAB
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.21
Holdup BI standard deviation: 0.09
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): 2.44
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: AKAP13
Protein A fragment: 1221-1230
Protein A site: 14-3-3_pSer1226
Protein A modification: (SEP)1226
Protein A sequence: SGRERSTPSL
Protein A construct: SGRER(pSer)TPSL (crude synthetic peptide)
Protein B protein: YWHAB
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.13
Holdup BI standard deviation: 0.06
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): 2.35
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: AKAP13
Protein A fragment: 2694-2703
Protein A site: 14-3-3_pSer2699
Protein A modification: (SEP)2699
Protein A sequence: LMRIPSFFPS
Protein A construct: LMRIP(pSer)FFPS (crude synthetic peptide)
Protein B protein: YWHAB
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.48
Holdup BI standard deviation: 0.26
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: 3.83
Experiment series 22
Protein A protein: AKAP13
Protein A fragment: 1890-1899
Protein A site: 14-3-3_pSer1895
Protein A modification: (SEP)1895
Protein A sequence: FANRRSQQSV
Protein A construct: FANRR(pSer)QQSV (crude synthetic peptide)
Protein B protein: YWHAB
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: -0.22
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): 1.53
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 23
Protein A protein: AKAP13
Protein A fragment: 1927-1936
Protein A site: 14-3-3_pSer1932
Protein A modification: (SEP)1932
Protein A sequence: SHSTDSLNKI
Protein A construct: SHSTD(pSer)LNKI (crude synthetic peptide)
Protein B protein: YWHAB
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.15
Holdup BI standard deviation: 0.11
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): 1.49
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: AKAP13
Protein A fragment: 1560-1569
Protein A site: 14-3-3_pSer1565
Protein A modification: (SEP)1565
Protein A sequence: PFRRHSWGPG
Protein A construct: PFRRH(pSer)WGPG (crude synthetic peptide)
Protein B protein: YWHAB
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.93
Holdup BI standard deviation: 0.06
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): 1.36
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: 5.01
Experiment series 25
Protein A protein: AKAP13
Protein A fragment: 2468-2477
Protein A site: 14-3-3_pSer2473
Protein A modification: (SEP)2473
Protein A sequence: FGGFDSHQMN
Protein A construct: FGGFD(pSer)HQMN (crude synthetic peptide)
Protein B protein: YWHAB
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.25
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): 1.01
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 26
Protein A protein: AKAP13
Protein A fragment: 2468-2477
Protein A site: 14-3-3_pSer2473
Protein A modification: (SEP)2473
Protein A sequence: FGGFDSHQMN
Protein A construct: FGGFD(pSer)HQMN (crude synthetic peptide)
Protein B protein: YWHAB
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.1
Holdup BI standard deviation: 0.17
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): 0.59
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 27
Protein A protein: AKAP13
Protein A fragment: 1890-1899
Protein A site: 14-3-3_pSer1895
Protein A modification: (SEP)1895
Protein A sequence: FANRRSQQSV
Protein A construct: FANRR(pSer)QQSV (crude synthetic peptide)
Protein B protein: YWHAB
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.1
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): 0.5
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
The affinity was below the detection threshold of the assay.
Experiment series 28
Protein A protein: AKAP13
Protein A fragment: 2694-2703
Protein A site: 14-3-3_pSer2699
Protein A modification: (SEP)2699
Protein A sequence: LMRIPSFFPS
Protein A construct: LMRIP(pSer)FFPS (crude synthetic peptide)
Protein B protein: YWHAB
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: -0.4
Holdup BI standard deviation: 1.25
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): 0.31
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 29
Protein A protein: AKAP13
Protein A fragment: 1640-1649
Protein A site: 14-3-3_pSer1645
Protein A modification: (SEP)1645
Protein A sequence: VDSLVSLSEE
Protein A construct: VDSLV(pSer)LSEE (crude synthetic peptide)
Protein B protein: YWHAB
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: -0.08
Holdup BI standard deviation: 0.33
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): 0.21
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 30
Protein A protein: AKAP13
Protein A fragment: 1637-1646
Protein A site: 14-3-3_pSer1642
Protein A modification: (SEP)1642
Protein A sequence: EERVDSLVSL
Protein A construct: EERVD(pSer)LVSL (crude synthetic peptide)
Protein B protein: YWHAB
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: -0.04
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): 0.2
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
The affinity was below the detection threshold of the assay.
Experiment series 31
Protein A protein: AKAP13
Protein A fragment: 1927-1936
Protein A site: 14-3-3_pSer1932
Protein A modification: (SEP)1932
Protein A sequence: SHSTDSLNKI
Protein A construct: SHSTD(pSer)LNKI (crude synthetic peptide)
Protein B protein: YWHAB
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.04
Holdup BI standard deviation: 0.19
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): 0.15
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 32
Protein A protein: AKAP13
Protein A fragment: 1637-1646
Protein A site: 14-3-3_pSer1642
Protein A modification: (SEP)1642
Protein A sequence: EERVDSLVSL
Protein A construct: EERVD(pSer)LVSL (crude synthetic peptide)
Protein B protein: YWHAB
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: -0.02
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): 0.1
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
The affinity was below the detection threshold of the assay.
Experiment series 33
Protein A protein: AKAP13
Protein A fragment: 1640-1649
Protein A site: 14-3-3_pSer1645
Protein A modification: (SEP)1645
Protein A sequence: VDSLVSLSEE
Protein A construct: VDSLV(pSer)LSEE (crude synthetic peptide)
Protein B protein: YWHAB
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.01
Holdup BI standard deviation: 0.06
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): 0.07
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 34
Protein A protein: AKAP13
Protein A fragment: 2406-2415
Protein A site: 14-3-3_pThr2411
Protein A modification: (TPO)2411
Protein A sequence: LFRSNTEEAL
Protein A construct: biotin-ttds-LFRSN(pThr)EEAL (HPLC-purified synthetic peptide)
Protein B protein: YWHAB
Protein B fragment: 1-246
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3beta
Measured dissociation constant (10-6M): 698
Standard deviation of dissociation constant (10-6M): 343
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: 3.16