Interaction Details


Details of experiments between P61981 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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

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: 0.16
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 134
P value: 3.29
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
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 4))

Experiment series 2

Protein A

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

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: 0.16
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 129
P value: 3.92
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: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

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: 0.14
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 115
P value: 3.28
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
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 4))

Experiment series 4

Protein A

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

Protein B

Protein: AKAP13
Fragment: 1224-1233
Site: 14-3-3_pSer1229
Modification: (SEP)1229
Sequence: ERSTPSLPCM
Construct: ERSTP(pSer)LPCM (crude synthetic peptide)
Average holdup BI: -0.1
Holdup BI standard deviation: 1.52
Immobilized partner concentration (10-6M): 129
P value: 0.05
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: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

Protein B

Protein: AKAP13
Fragment: 1224-1233
Site: 14-3-3_pSer1229
Modification: (SEP)1229
Sequence: ERSTPSLPCM
Construct: ERSTP(pSer)LPCM (crude synthetic peptide)
Average holdup BI: -0.07
Holdup BI standard deviation: 1.37
Immobilized partner concentration (10-6M): 115
P value: 0.04
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
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 4))

Experiment series 6

Protein A

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

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: 0.35
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 129
P value: 4.85
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.62
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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

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: 0.33
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 134
P value: 6.03
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
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 4))

Experiment series 8

Protein A

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

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: 0.29
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 115
P value: 6.31
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.57
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 9

Protein A

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

Protein B

Protein: AKAP13
Fragment: 1560-1569
Site: 14-3-3_pSer1565
Modification: (SEP)1565
Sequence: PFRRHSWGPG
Construct: PFRRH(pSer)WGPG (crude synthetic peptide)
Average holdup BI: -0.14
Holdup BI standard deviation: 1.24
Immobilized partner concentration (10-6M): 129
P value: 0.09
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 10

Protein A

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

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): 129
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 11

Protein A

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

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: -0.31
Holdup BI standard deviation: 0.3
Immobilized partner concentration (10-6M): 134
P value: 1.12
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
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 4))

Experiment series 12

Protein A

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

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: -0.08
Holdup BI standard deviation: 0.16
Immobilized partner concentration (10-6M): 129
P value: 0.47
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 13

Protein A

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

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: 0.07
Holdup BI standard deviation: 0.13
Immobilized partner concentration (10-6M): 115
P value: 0.51
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
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 4))

Experiment series 14

Protein A

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

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: 0.09
Holdup BI standard deviation: 0.11
Immobilized partner concentration (10-6M): 134
P value: 0.81
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
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 4))

Experiment series 15

Protein A

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

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: -0.17
Holdup BI standard deviation: 0.25
Immobilized partner concentration (10-6M): 129
P value: 0.95
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: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

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: 0.07
Holdup BI standard deviation: 0.1
Immobilized partner concentration (10-6M): 115
P value: 0.65
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
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 4))

Experiment series 17

Protein A

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

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: 0.4
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 134
P value: 4.17
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.7
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 18

Protein A

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

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: 0.32
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 129
P value: 5.19
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.57
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: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

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: 0.3
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 115
P value: 3.78
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.6
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 20

Protein A

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

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: 0.75
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 129
P value: 5.65
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.38
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 21

Protein A

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

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: 0.72
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 134
P value: 5.66
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.29
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 22

Protein A

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

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: 0.67
Holdup BI standard deviation: 0.1
Immobilized partner concentration (10-6M): 115
P value: 2.84
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.27
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 23

Protein A

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

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: 0.03
Holdup BI standard deviation: 0.27
Immobilized partner concentration (10-6M): 134
P value: 0.07
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
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 4))

Experiment series 24

Protein A

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

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: 0.26
Holdup BI standard deviation: 0.2
Immobilized partner concentration (10-6M): 129
P value: 1.38
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 25

Protein A

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

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: 0.2
Holdup BI standard deviation: 0.24
Immobilized partner concentration (10-6M): 115
P value: 0.84
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
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 4))

Experiment series 26

Protein A

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

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: 0.71
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 129
P value: 6.93
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.28
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 27

Protein A

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

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: 0.65
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 134
P value: 5.63
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.15
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 28

Protein A

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

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: 0.66
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 115
P value: 6.38
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.25
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 29

Protein A

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

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: 0.16
Holdup BI standard deviation: 0.29
Immobilized partner concentration (10-6M): 134
P value: 0.49
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
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 4))

Experiment series 30

Protein A

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

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: 0.21
Holdup BI standard deviation: 0.43
Immobilized partner concentration (10-6M): 129
P value: 0.44
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 31

Protein A

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

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: -0.3
Holdup BI standard deviation: 0.53
Immobilized partner concentration (10-6M): 115
P value: 0.64
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
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 4))

Experiment series 32

Protein A

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

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: 0.14
Holdup BI standard deviation: 0.27
Immobilized partner concentration (10-6M): 134
P value: 0.44
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
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 4))

Experiment series 33

Protein A

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

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: -0.05
Holdup BI standard deviation: 0.09
Immobilized partner concentration (10-6M): 129
P value: 0.65
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 34

Protein A

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

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: -0.2
Holdup BI standard deviation: 0.7
Immobilized partner concentration (10-6M): 115
P value: 0.23
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
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 4))

Experiment series 35

Protein A

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

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: 0.52
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 129
P value: 6.26
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.93
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 36

Protein A

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

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: 0.52
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 134
P value: 7.2
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.91
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 37

Protein A

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

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: 0.63
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 115
P value: 5.52
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.19
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 38

Protein A

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

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: 0.38
Holdup BI standard deviation: 0.08
Immobilized partner concentration (10-6M): 134
P value: 3.98
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.66
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 39

Protein A

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

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: 0.36
Holdup BI standard deviation: 0.18
Immobilized partner concentration (10-6M): 129
P value: 1.73
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.64
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 40

Protein A

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

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: 0.29
Holdup BI standard deviation: 0.12
Immobilized partner concentration (10-6M): 115
P value: 2.28
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.57
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 41

Protein A

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

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: 0.22
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 134
P value: 5.27
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
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 4))

Experiment series 42

Protein A

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

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: 0.26
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 129
P value: 4.53
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 43

Protein A

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

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: 0.22
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 115
P value: 3.4
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
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 4))

Experiment series 44

Protein A

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

Protein B

Protein: AKAP13
Fragment: 2694-2703
Site: 14-3-3_pSer2699
Modification: (SEP)2699
Sequence: LMRIPSFFPS
Construct: LMRIP(pSer)FFPS (crude synthetic peptide)
Average holdup BI: -0.48
Holdup BI standard deviation: 1.41
Immobilized partner concentration (10-6M): 129
P value: 0.33
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 45

Protein A

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

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: 0.31
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 129
P value: 7.04
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.55
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 46

Protein A

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

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): 300
Standard deviation of dissociation constant (10-6M): 12
Experiment method: competitive Fluorescence Polarization
Details of affinity fitting: Competitive binding equation in ProFit
Date of measurement: 2022-01-10
Number of measurements: 3
Measured pKd: 3.52
DOI: 10.1101/2022.12.08.519103

Experiment series 47

Protein A

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

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): 394
Standard deviation of dissociation constant (10-6M): 63
Experiment method: competitive Fluorescence Polarization
Details of affinity fitting: Competitive binding equation in ProFit
Date of measurement: 2022-01-18
Number of measurements: 3
Measured pKd: 3.4
DOI: 10.1101/2022.12.08.519103

Experiment series 48

Protein A

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

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: 0.3
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 134
P value: 4.98
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
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 4))

Experiment series 49

Protein A

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

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: 0.33
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 115
P value: 3.98
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.64
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 50

Protein A

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

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: 0.99
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 134
P value: 5.79
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 5.71
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 51

Protein A

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

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: 0.02
Immobilized partner concentration (10-6M): 129
P value: 7.49
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 5.39
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 52

Protein A

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

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): 115
P value: 5.57
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 5.43
DOI: 10.1101/2022.12.08.519103
Experimental Details

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