Interaction Details


Details of experiments between Q6IQ23 and P61981
Note that the results of all experiments are listed, regardless of the modification states of the fragments.


Experiment series 1

Protein A

Protein: PLEKHA7
Fragment: 1058-1067
Site: 14-3-3_pSer1063
Modification: (SEP)1063
Sequence: LERLYSGDHQ
Construct: LERLY(pSer)GDHQ (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.04
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 134
P value: 0.82
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: PLEKHA7
Fragment: 1058-1067
Site: 14-3-3_pSer1063
Modification: (SEP)1063
Sequence: LERLYSGDHQ
Construct: LERLY(pSer)GDHQ (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 129
P value: 0.01
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: PLEKHA7
Fragment: 1105-1114
Site: 14-3-3_pSer1110
Modification: (SEP)1110
Sequence: SSRYLSRPLP
Construct: SSRYL(pSer)RPLP (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.42
Holdup BI standard deviation: 0.12
Immobilized partner concentration (10-6M): 134
P value: 2.33
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.73
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: PLEKHA7
Fragment: 1105-1114
Site: 14-3-3_pSer1110
Modification: (SEP)1110
Sequence: SSRYLSRPLP
Construct: SSRYL(pSer)RPLP (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.36
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 129
P value: 6.21
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 5

Protein A

Protein: PLEKHA7
Fragment: 443-452
Site: 14-3-3_pSer448
Modification: (SEP)448
Sequence: KGDSRSLPLD
Construct: KGDSR(pSer)LPLD (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.92
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 129
P value: 9
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.92
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: PLEKHA7
Fragment: 443-452
Site: 14-3-3_pSer448
Modification: (SEP)448
Sequence: KGDSRSLPLD
Construct: KGDSR(pSer)LPLD (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.9
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 134
P value: 10.43
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.82
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 7

Protein A

Protein: PLEKHA7
Fragment: 443-452
Site: 14-3-3_pSer448
Modification: (SEP)448
Sequence: KGDSRSLPLD
Construct: biotin-ttds-KGDSR(pSer)LPLD (HPLC-purified synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Measured dissociation constant (10-6M): 28
Standard deviation of dissociation constant (10-6M): 3.6
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: 4.55
DOI: 10.1101/2022.12.08.519103

Experiment series 8

Protein A

Protein: PLEKHA7
Fragment: 560-569
Site: 14-3-3_pSer565
Modification: (SEP)565
Sequence: VPRSISVPPS
Construct: VPRSI(pSer)VPPS (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.93
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 129
P value: 8.79
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 5.01
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: PLEKHA7
Fragment: 560-569
Site: 14-3-3_pSer565
Modification: (SEP)565
Sequence: VPRSISVPPS
Construct: VPRSI(pSer)VPPS (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.92
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 134
P value: 4.61
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.94
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 10

Protein A

Protein: PLEKHA7
Fragment: 599-608
Site: 14-3-3_pSer604
Modification: (SEP)604
Sequence: PDQRRSVDIS
Construct: PDQRR(pSer)VDIS (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.08
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 134
P value: 1.29
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 11

Protein A

Protein: PLEKHA7
Fragment: 599-608
Site: 14-3-3_pSer604
Modification: (SEP)604
Sequence: PDQRRSVDIS
Construct: PDQRR(pSer)VDIS (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.1
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 129
P value: 2.35
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 12

Protein A

Protein: PLEKHA7
Fragment: 603-612
Site: 14-3-3_pSer608
Modification: (SEP)608
Sequence: RSVDISLGDS
Construct: RSVDI(pSer)LGDS (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: -0.12
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 134
P value: 2.27
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 13

Protein A

Protein: PLEKHA7
Fragment: 603-612
Site: 14-3-3_pSer608
Modification: (SEP)608
Sequence: RSVDISLGDS
Construct: RSVDI(pSer)LGDS (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: -0.05
Holdup BI standard deviation: 0.1
Immobilized partner concentration (10-6M): 129
P value: 0.51
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: PLEKHA7
Fragment: 626-635
Site: 14-3-3_pSer631
Modification: (SEP)631
Sequence: HVDRRSMPSM
Construct: HVDRR(pSer)MPSM (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.42
Holdup BI standard deviation: 0.13
Immobilized partner concentration (10-6M): 129
P value: 3.29
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.75
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: PLEKHA7
Fragment: 981-990
Site: 14-3-3_pSer986
Modification: (SEP)986
Sequence: LRSYVSEPEL
Construct: LRSYV(pSer)EPEL (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.77
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 129
P value: 5.3
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.42
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: PLEKHA7
Fragment: 981-990
Site: 14-3-3_pSer986
Modification: (SEP)986
Sequence: LRSYVSEPEL
Construct: LRSYV(pSer)EPEL (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.66
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 134
P value: 5.42
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.16
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: PLEKHA7
Fragment: 1008-1017
Site: 14-3-3_pThr1013
Modification: (TPO)1013
Sequence: ESRYQTLPGR
Construct: ESRYQ(pThr)LPGR (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.83
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 129
P value: 8.4
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.57
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 18

Protein A

Protein: PLEKHA7
Fragment: 1008-1017
Site: 14-3-3_pThr1013
Modification: (TPO)1013
Sequence: ESRYQTLPGR
Construct: ESRYQ(pThr)LPGR (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.82
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 134
P value: 8.09
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.52
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 19

Protein A

Protein: PLEKHA7
Fragment: 1032-1041
Site: 14-3-3_pThr1037
Modification: (TPO)1037
Sequence: IAPYVTLRRG
Construct: IAPYV(pThr)LRRG (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.42
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 134
P value: 3.81
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.72
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: PLEKHA7
Fragment: 1032-1041
Site: 14-3-3_pThr1037
Modification: (TPO)1037
Sequence: IAPYVTLRRG
Construct: IAPYV(pThr)LRRG (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.38
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 129
P value: 5.21
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.68
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: PLEKHA7
Fragment: 467-476
Site: 14-3-3_pThr472
Modification: (TPO)472
Sequence: PENYQTLPKS
Construct: PENYQ(pThr)LPKS (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.29
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 129
P value: 8.12
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.5
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 22

Protein A

Protein: PLEKHA7
Fragment: 467-476
Site: 14-3-3_pThr472
Modification: (TPO)472
Sequence: PENYQTLPKS
Construct: PENYQ(pThr)LPKS (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.29
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 134
P value: 6.08
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 23

Protein A

Protein: PLEKHA7
Fragment: 533-542
Site: 14-3-3_pThr538
Modification: (TPO)538
Sequence: RHGSPTAPI-
Construct: FRHGSP(pThr)API (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.87
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 129
P value: 2.32
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.7
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 24

Protein A

Protein: PLEKHA7
Fragment: 533-542
Site: 14-3-3_pThr538
Modification: (TPO)538
Sequence: RHGSPTAPI-
Construct: FRHGSP(pThr)API (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.85
Holdup BI standard deviation: 0.08
Immobilized partner concentration (10-6M): 134
P value: 4.55
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.63
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 25

Protein A

Protein: PLEKHA7
Fragment: 882-891
Site: 14-3-3_pThr887
Modification: (TPO)887
Sequence: FPQLQTYVPY
Construct: FPQLQ(pThr)YVPY (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: -0.09
Holdup BI standard deviation: 0.98
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 26

Protein A

Protein: PLEKHA7
Fragment: 882-891
Site: 14-3-3_pThr887
Modification: (TPO)887
Sequence: FPQLQTYVPY
Construct: FPQLQ(pThr)YVPY (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: -0.17
Holdup BI standard deviation: 1.1
Immobilized partner concentration (10-6M): 129
P value: 0.12
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 27

Protein A

Protein: PLEKHA7
Fragment: 897-906
Site: 14-3-3_pThr902
Modification: (TPO)902
Sequence: QLRKVTSPLQ
Construct: QLRKV(pThr)SPLQ (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.25
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 134
P value: 3.68
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 28

Protein A

Protein: PLEKHA7
Fragment: 897-906
Site: 14-3-3_pThr902
Modification: (TPO)902
Sequence: QLRKVTSPLQ
Construct: QLRKV(pThr)SPLQ (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.22
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 129
P value: 3.96
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 29

Protein A

Protein: PLEKHA7
Fragment: 1058-1067
Site: 14-3-3_pSer1063
Modification: (SEP)1063
Sequence: LERLYSGDHQ
Construct: LERLY(pSer)GDHQ (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.06
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 115
P value: 0.76
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: PLEKHA7
Fragment: 1105-1114
Site: 14-3-3_pSer1110
Modification: (SEP)1110
Sequence: SSRYLSRPLP
Construct: SSRYL(pSer)RPLP (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.38
Holdup BI standard deviation: 0.08
Immobilized partner concentration (10-6M): 115
P value: 3.15
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.73
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 31

Protein A

Protein: PLEKHA7
Fragment: 443-452
Site: 14-3-3_pSer448
Modification: (SEP)448
Sequence: KGDSRSLPLD
Construct: KGDSR(pSer)LPLD (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.86
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 115
P value: 8.68
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.75
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: PLEKHA7
Fragment: 560-569
Site: 14-3-3_pSer565
Modification: (SEP)565
Sequence: VPRSISVPPS
Construct: VPRSI(pSer)VPPS (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.99
Holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 115
P value: 8.22
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 5.91
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: PLEKHA7
Fragment: 599-608
Site: 14-3-3_pSer604
Modification: (SEP)604
Sequence: PDQRRSVDIS
Construct: PDQRR(pSer)VDIS (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.18
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 115
P value: 2.73
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 34

Protein A

Protein: PLEKHA7
Fragment: 603-612
Site: 14-3-3_pSer608
Modification: (SEP)608
Sequence: RSVDISLGDS
Construct: RSVDI(pSer)LGDS (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.02
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 115
P value: 0.31
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: PLEKHA7
Fragment: 626-635
Site: 14-3-3_pSer631
Modification: (SEP)631
Sequence: HVDRRSMPSM
Construct: HVDRR(pSer)MPSM (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: -0.26
Holdup BI standard deviation: 1.3
Immobilized partner concentration (10-6M): 115
P value: 0.16
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 36

Protein A

Protein: PLEKHA7
Fragment: 981-990
Site: 14-3-3_pSer986
Modification: (SEP)986
Sequence: LRSYVSEPEL
Construct: LRSYV(pSer)EPEL (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.72
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 115
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
Measured pKd: 4.36
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: PLEKHA7
Fragment: 1008-1017
Site: 14-3-3_pThr1013
Modification: (TPO)1013
Sequence: ESRYQTLPGR
Construct: ESRYQ(pThr)LPGR (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.84
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 115
P value: 6.78
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.67
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: PLEKHA7
Fragment: 1032-1041
Site: 14-3-3_pThr1037
Modification: (TPO)1037
Sequence: IAPYVTLRRG
Construct: IAPYV(pThr)LRRG (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.37
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 115
P value: 4.85
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.73
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: PLEKHA7
Fragment: 467-476
Site: 14-3-3_pThr472
Modification: (TPO)472
Sequence: PENYQTLPKS
Construct: PENYQ(pThr)LPKS (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.44
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 115
P value: 6.81
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.85
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 40

Protein A

Protein: PLEKHA7
Fragment: 533-542
Site: 14-3-3_pThr538
Modification: (TPO)538
Sequence: RHGSPTAPI-
Construct: FRHGSP(pThr)API (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.94
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 115
P value: 5.1
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 5.14
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: PLEKHA7
Fragment: 882-891
Site: 14-3-3_pThr887
Modification: (TPO)887
Sequence: FPQLQTYVPY
Construct: FPQLQ(pThr)YVPY (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.31
Holdup BI standard deviation: 0.1
Immobilized partner concentration (10-6M): 115
P value: 2.45
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 42

Protein A

Protein: PLEKHA7
Fragment: 897-906
Site: 14-3-3_pThr902
Modification: (TPO)902
Sequence: QLRKVTSPLQ
Construct: QLRKV(pThr)SPLQ (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.23
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 115
P value: 3.39
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))