Interaction Details


Details of experiments between P63104 and Q12802
Note that the results of all experiments are listed, regardless of the modification states of the fragments.


Experiment series 1

Protein A

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta

Protein B

Protein: AKAP13
Fragment: 1221-1230
Site: 14-3-3_pSer1226
Modification: (SEP)1226
Sequence: SGRERSTPSL
Construct: SGRER(pSer)TPSL (crude synthetic peptide)
Average holdup BI: 0.11
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 132
P value: 1.88
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 2

Protein A

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta

Protein B

Protein: AKAP13
Fragment: 1502-1511
Site: 14-3-3_pSer1507
Modification: (SEP)1507
Sequence: SRDRQSLDGF
Construct: SRDRQ(pSer)LDGF (crude synthetic peptide)
Average holdup BI: 0.21
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 132
P value: 3.88
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 3

Protein A

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta

Protein B

Protein: AKAP13
Fragment: 1597-1606
Site: 14-3-3_pSer1602
Modification: (SEP)1602
Sequence: HRRSFSLEGL
Construct: HRRSF(pSer)LEGL (crude synthetic peptide)
Measured dissociation constant (10-6M): <100
Immobilized partner concentration (10-6M): 132
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 1
Measured pKd: 4
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 4

Protein A

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta

Protein B

Protein: AKAP13
Fragment: 1637-1646
Site: 14-3-3_pSer1642
Modification: (SEP)1642
Sequence: EERVDSLVSL
Construct: EERVD(pSer)LVSL (crude synthetic peptide)
Average holdup BI: -0.14
Holdup BI standard deviation: 0.13
Immobilized partner concentration (10-6M): 132
P value: 1.41
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 5

Protein A

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta

Protein B

Protein: AKAP13
Fragment: 1640-1649
Site: 14-3-3_pSer1645
Modification: (SEP)1645
Sequence: VDSLVSLSEE
Construct: VDSLV(pSer)LSEE (crude synthetic peptide)
Average holdup BI: -0.16
Holdup BI standard deviation: 0.24
Immobilized partner concentration (10-6M): 132
P value: 0.85
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 6

Protein A

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta

Protein B

Protein: AKAP13
Fragment: 1745-1754
Site: 14-3-3_pSer1750
Modification: (SEP)1750
Sequence: VSRTFSYIKN
Construct: VSRTF(pSer)YIKN (crude synthetic peptide)
Average holdup BI: 0.18
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 132
P value: 3.97
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 7

Protein A

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta

Protein B

Protein: AKAP13
Fragment: 1871-1880
Site: 14-3-3_pSer1876
Modification: (SEP)1876
Sequence: KERPRSAVLL
Construct: KERPR(pSer)AVLL (crude synthetic peptide)
Average holdup BI: 0.62
Holdup BI standard deviation: 0.11
Immobilized partner concentration (10-6M): 132
P value: 4.25
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.08
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 8

Protein A

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta

Protein B

Protein: AKAP13
Fragment: 1890-1899
Site: 14-3-3_pSer1895
Modification: (SEP)1895
Sequence: FANRRSQQSV
Construct: FANRR(pSer)QQSV (crude synthetic peptide)
Average holdup BI: 0.08
Holdup BI standard deviation: 0.09
Immobilized partner concentration (10-6M): 132
P value: 0.91
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 9

Protein A

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta

Protein B

Protein: AKAP13
Fragment: 1924-1933
Site: 14-3-3_pSer1929
Modification: (SEP)1929
Sequence: KFLSHSTDSL
Construct: KFLSH(pSer)TDSL (crude synthetic peptide)
Average holdup BI: 0.52
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 132
P value: 5.76
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.91
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 10

Protein A

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta

Protein B

Protein: AKAP13
Fragment: 1927-1936
Site: 14-3-3_pSer1932
Modification: (SEP)1932
Sequence: SHSTDSLNKI
Construct: SHSTD(pSer)LNKI (crude synthetic peptide)
Average holdup BI: 0.09
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 132
P value: 1.53
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 11

Protein A

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta

Protein B

Protein: AKAP13
Fragment: 2468-2477
Site: 14-3-3_pSer2473
Modification: (SEP)2473
Sequence: FGGFDSHQMN
Construct: FGGFD(pSer)HQMN (crude synthetic peptide)
Average holdup BI: 0
Holdup BI standard deviation: 0.11
Immobilized partner concentration (10-6M): 132
P value: 0.02
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 12

Protein A

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta

Protein B

Protein: AKAP13
Fragment: 2493-2502
Site: 14-3-3_pSer2498
Modification: (SEP)2498
Sequence: LRRTESDSGL
Construct: LRRTE(pSer)DSGL (crude synthetic peptide)
Average holdup BI: 0.38
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 132
P value: 5.37
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.66
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 13

Protein A

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta

Protein B

Protein: AKAP13
Fragment: 2558-2567
Site: 14-3-3_pSer2563
Modification: (SEP)2563
Sequence: LTRSLSRPSS
Construct: LTRSL(pSer)RPSS (crude synthetic peptide)
Average holdup BI: 0.2
Holdup BI standard deviation: 0.22
Immobilized partner concentration (10-6M): 132
P value: 0.82
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 14

Protein A

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta

Protein B

Protein: AKAP13
Fragment: 2562-2571
Site: 14-3-3_pSer2567
Modification: (SEP)2567
Sequence: LSRPSSLIEQ
Construct: LSRPS(pSer)LIEQ (crude synthetic peptide)
Average holdup BI: 0.17
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 132
P value: 3.28
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 15

Protein A

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta

Protein B

Protein: AKAP13
Fragment: 2728-2737
Site: 14-3-3_pSer2733
Modification: (SEP)2733
Sequence: SPKRNSISRT
Construct: SPKRN(pSer)ISRT (crude synthetic peptide)
Average holdup BI: 0.19
Holdup BI standard deviation: 0.16
Immobilized partner concentration (10-6M): 132
P value: 1
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 16

Protein A

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta

Protein B

Protein: AKAP13
Fragment: 2406-2415
Site: 14-3-3_pThr2411
Modification: (TPO)2411
Sequence: LFRSNTEEAL
Construct: biotin-ttds-LFRSN(pThr)EEAL (HPLC-purified synthetic peptide)
Measured dissociation constant (10-6M): 763
Standard deviation of dissociation constant (10-6M): 76
Experiment method: competitive Fluorescence Polarization
Details of affinity fitting: Competitive binding equation in ProFit
Date of measurement: 2022-01-19
Number of measurements: 3
Measured pKd: 3.11
DOI: 10.1101/2022.12.08.519103

Experiment series 17

Protein A

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta

Protein B

Protein: AKAP13
Fragment: 2406-2415
Site: 14-3-3_pThr2411
Modification: (TPO)2411
Sequence: LFRSNTEEAL
Construct: biotin-ttds-LFRSN(pThr)EEAL (HPLC-purified synthetic peptide)
Measured dissociation constant (10-6M): 1213
Standard deviation of dissociation constant (10-6M): 200
Experiment method: competitive Fluorescence Polarization
Details of affinity fitting: Competitive binding equation in ProFit
Date of measurement: 2022-02-10
Number of measurements: 3
Measured pKd: 2.92
DOI: 10.1101/2022.12.08.519103

Experiment series 18

Protein A

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta

Protein B

Protein: AKAP13
Fragment: 2406-2415
Site: 14-3-3_pThr2411
Modification: (TPO)2411
Sequence: LFRSNTEEAL
Construct: LFRSN(pThr)EEAL (crude synthetic peptide)
Average holdup BI: 0.17
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 132
P value: 4.35
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))

Experiment series 19

Protein A

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta

Protein B

Protein: AKAP13
Fragment: 2462-2471
Site: 14-3-3_pThr2467
Modification: (TPO)2467
Sequence: PRRAETFGGF
Construct: PRRAE(pThr)FGGF (crude synthetic peptide)
Average holdup BI: 0.97
Holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 132
P value: 7.38
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 5.33
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))