Interaction Details


Details of experiments between P62258 and Q6IQ23
Note that the results of all experiments are listed, regardless of the modification states of the fragments.


Experiment series 1

Protein A

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Protein B

Protein: PLEKHA7
Fragment: 1058-1067
Site: 14-3-3_pSer1063
Modification: (SEP)1063
Sequence: LERLYSGDHQ
Construct: LERLY(pSer)GDHQ (crude synthetic peptide)
Average holdup BI: 0.03
Holdup BI standard deviation: 0.11
Immobilized partner concentration (10-6M): 116
P value: 0.22
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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Protein B

Protein: PLEKHA7
Fragment: 1058-1067
Site: 14-3-3_pSer1063
Modification: (SEP)1063
Sequence: LERLYSGDHQ
Construct: LERLY(pSer)GDHQ (crude synthetic peptide)
Average holdup BI: -0.02
Holdup BI standard deviation: 0.08
Immobilized partner concentration (10-6M): 143
P value: 0.17
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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Protein B

Protein: PLEKHA7
Fragment: 1105-1114
Site: 14-3-3_pSer1110
Modification: (SEP)1110
Sequence: SSRYLSRPLP
Construct: SSRYL(pSer)RPLP (crude synthetic peptide)
Average holdup BI: 0.07
Holdup BI standard deviation: 0.17
Immobilized partner concentration (10-6M): 116
P value: 0.29
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 4

Protein A

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Protein B

Protein: PLEKHA7
Fragment: 1105-1114
Site: 14-3-3_pSer1110
Modification: (SEP)1110
Sequence: SSRYLSRPLP
Construct: SSRYL(pSer)RPLP (crude synthetic peptide)
Average holdup BI: 0.04
Holdup BI standard deviation: 0.13
Immobilized partner concentration (10-6M): 143
P value: 0.28
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 5

Protein A

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Protein B

Protein: PLEKHA7
Fragment: 421-430
Site: 14-3-3_pSer426
Modification: (SEP)426
Sequence: NPEKHSQRKS
Construct: NPEKH(pSer)QRKS (crude synthetic peptide)
Average holdup BI: -0.21
Holdup BI standard deviation: 0.99
Immobilized partner concentration (10-6M): 116
P value: 0.1
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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Protein B

Protein: PLEKHA7
Fragment: 443-452
Site: 14-3-3_pSer448
Modification: (SEP)448
Sequence: KGDSRSLPLD
Construct: KGDSR(pSer)LPLD (crude synthetic peptide)
Average holdup BI: 0.64
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 116
P value: 9.03
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.18
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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Protein B

Protein: PLEKHA7
Fragment: 443-452
Site: 14-3-3_pSer448
Modification: (SEP)448
Sequence: KGDSRSLPLD
Construct: KGDSR(pSer)LPLD (crude synthetic peptide)
Average holdup BI: 0.64
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 143
P value: 7.61
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.1
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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Protein B

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)
Measured dissociation constant (10-6M): 93
Standard deviation of dissociation constant (10-6M): 11
Experiment method: competitive Fluorescence Polarization
Details of affinity fitting: Competitive binding equation in ProFit
Date of measurement: 2022-02-09
Number of measurements: 3
Measured pKd: 4.03
DOI: 10.1101/2022.12.08.519103

Experiment series 9

Protein A

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Protein B

Protein: PLEKHA7
Fragment: 560-569
Site: 14-3-3_pSer565
Modification: (SEP)565
Sequence: VPRSISVPPS
Construct: VPRSI(pSer)VPPS (crude synthetic peptide)
Average holdup BI: 0.71
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 116
P value: 3.93
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.32
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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Protein B

Protein: PLEKHA7
Fragment: 560-569
Site: 14-3-3_pSer565
Modification: (SEP)565
Sequence: VPRSISVPPS
Construct: VPRSI(pSer)VPPS (crude synthetic peptide)
Average holdup BI: 0.73
Holdup BI standard deviation: 0.11
Immobilized partner concentration (10-6M): 143
P value: 5.25
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.28
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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Protein B

Protein: PLEKHA7
Fragment: 599-608
Site: 14-3-3_pSer604
Modification: (SEP)604
Sequence: PDQRRSVDIS
Construct: PDQRR(pSer)VDIS (crude synthetic peptide)
Average holdup BI: 0.07
Holdup BI standard deviation: 0.08
Immobilized partner concentration (10-6M): 116
P value: 0.98
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 12

Protein A

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Protein B

Protein: PLEKHA7
Fragment: 599-608
Site: 14-3-3_pSer604
Modification: (SEP)604
Sequence: PDQRRSVDIS
Construct: PDQRR(pSer)VDIS (crude synthetic peptide)
Average holdup BI: 0.06
Holdup BI standard deviation: 0.11
Immobilized partner concentration (10-6M): 143
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 13

Protein A

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Protein B

Protein: PLEKHA7
Fragment: 603-612
Site: 14-3-3_pSer608
Modification: (SEP)608
Sequence: RSVDISLGDS
Construct: RSVDI(pSer)LGDS (crude synthetic peptide)
Average holdup BI: -0.06
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 116
P value: 1.57
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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Protein B

Protein: PLEKHA7
Fragment: 603-612
Site: 14-3-3_pSer608
Modification: (SEP)608
Sequence: RSVDISLGDS
Construct: RSVDI(pSer)LGDS (crude synthetic peptide)
Average holdup BI: -0.09
Holdup BI standard deviation: 0.08
Immobilized partner concentration (10-6M): 143
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 15

Protein A

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Protein B

Protein: PLEKHA7
Fragment: 981-990
Site: 14-3-3_pSer986
Modification: (SEP)986
Sequence: LRSYVSEPEL
Construct: LRSYV(pSer)EPEL (crude synthetic peptide)
Average holdup BI: 0.61
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 143
P value: 4.73
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.05
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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Protein B

Protein: PLEKHA7
Fragment: 981-990
Site: 14-3-3_pSer986
Modification: (SEP)986
Sequence: LRSYVSEPEL
Construct: LRSYV(pSer)EPEL (crude synthetic peptide)
Average holdup BI: 0.47
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 116
P value: 4.42
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.89
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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Protein B

Protein: PLEKHA7
Fragment: 1008-1017
Site: 14-3-3_pThr1013
Modification: (TPO)1013
Sequence: ESRYQTLPGR
Construct: ESRYQ(pThr)LPGR (crude synthetic peptide)
Average holdup BI: 0.64
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 116
P value: 7.74
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.18
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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Protein B

Protein: PLEKHA7
Fragment: 1008-1017
Site: 14-3-3_pThr1013
Modification: (TPO)1013
Sequence: ESRYQTLPGR
Construct: ESRYQ(pThr)LPGR (crude synthetic peptide)
Average holdup BI: 0.66
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 143
P value: 6.97
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.14
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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Protein B

Protein: PLEKHA7
Fragment: 1032-1041
Site: 14-3-3_pThr1037
Modification: (TPO)1037
Sequence: IAPYVTLRRG
Construct: IAPYV(pThr)LRRG (crude synthetic peptide)
Average holdup BI: 0.33
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 143
P value: 4.41
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 20

Protein A

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Protein B

Protein: PLEKHA7
Fragment: 1032-1041
Site: 14-3-3_pThr1037
Modification: (TPO)1037
Sequence: IAPYVTLRRG
Construct: IAPYV(pThr)LRRG (crude synthetic peptide)
Average holdup BI: 0.27
Holdup BI standard deviation: 0.09
Immobilized partner concentration (10-6M): 116
P value: 2.77
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.51
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 21

Protein A

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Protein B

Protein: PLEKHA7
Fragment: 467-476
Site: 14-3-3_pThr472
Modification: (TPO)472
Sequence: PENYQTLPKS
Construct: PENYQ(pThr)LPKS (crude synthetic peptide)
Average holdup BI: 0.17
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 143
P value: 4.29
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.15
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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Protein B

Protein: PLEKHA7
Fragment: 467-476
Site: 14-3-3_pThr472
Modification: (TPO)472
Sequence: PENYQTLPKS
Construct: PENYQ(pThr)LPKS (crude synthetic peptide)
Average holdup BI: 0.12
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 116
P value: 4.4
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 23

Protein A

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Protein B

Protein: PLEKHA7
Fragment: 533-542
Site: 14-3-3_pThr538
Modification: (TPO)538
Sequence: RHGSPTAPI-
Construct: FRHGSP(pThr)API (crude synthetic peptide)
Measured dissociation constant (10-6M): <10
Immobilized partner concentration (10-6M): 143
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 1
Measured pKd: 5
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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Protein B

Protein: PLEKHA7
Fragment: 533-542
Site: 14-3-3_pThr538
Modification: (TPO)538
Sequence: RHGSPTAPI-
Construct: FRHGSP(pThr)API (crude synthetic peptide)
Average holdup BI: 0.81
Holdup BI standard deviation: 0.11
Immobilized partner concentration (10-6M): 116
P value: 4.16
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.57
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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Protein B

Protein: PLEKHA7
Fragment: 882-891
Site: 14-3-3_pThr887
Modification: (TPO)887
Sequence: FPQLQTYVPY
Construct: FPQLQ(pThr)YVPY (crude synthetic peptide)
Average holdup BI: 0.31
Holdup BI standard deviation: 0.23
Immobilized partner concentration (10-6M): 116
P value: 1.4
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.59
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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Protein B

Protein: PLEKHA7
Fragment: 897-906
Site: 14-3-3_pThr902
Modification: (TPO)902
Sequence: QLRKVTSPLQ
Construct: QLRKV(pThr)SPLQ (crude synthetic peptide)
Average holdup BI: 0.08
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 116
P value: 1.34
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 27

Protein A

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon

Protein B

Protein: PLEKHA7
Fragment: 897-906
Site: 14-3-3_pThr902
Modification: (TPO)902
Sequence: QLRKVTSPLQ
Construct: QLRKV(pThr)SPLQ (crude synthetic peptide)
Average holdup BI: 0.12
Holdup BI standard deviation: 0.1
Immobilized partner concentration (10-6M): 143
P value: 1.31
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))