Interaction Details


Details of experiments between Q6IQ23 and P63104
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: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta
Average holdup BI: 0.01
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 132
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 2

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: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta
Average holdup BI: 0.21
Holdup BI standard deviation: 0.08
Immobilized partner concentration (10-6M): 132
P value: 2.77
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: 443-452
Site: 14-3-3_pSer448
Modification: (SEP)448
Sequence: KGDSRSLPLD
Construct: KGDSR(pSer)LPLD (crude synthetic peptide)

Protein B

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta
Average holdup BI: 0.81
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 132
P value: 8.24
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.5
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: 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: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta
Measured dissociation constant (10-6M): 98
Standard deviation of dissociation constant (10-6M): 4
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: 4.01
DOI: 10.1101/2022.12.08.519103

Experiment series 5

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: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta
Measured dissociation constant (10-6M): 100
Standard deviation of dissociation constant (10-6M): 5
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: 4
DOI: 10.1101/2022.12.08.519103

Experiment series 6

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: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta
Average holdup BI: 0.87
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 132
P value: 8.17
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.69
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: PLEKHA7
Fragment: 599-608
Site: 14-3-3_pSer604
Modification: (SEP)604
Sequence: PDQRRSVDIS
Construct: PDQRR(pSer)VDIS (crude synthetic peptide)

Protein B

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta
Average holdup BI: 0.05
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 132
P value: 1.13
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 8

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: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta
Average holdup BI: -0.06
Holdup BI standard deviation: 0.09
Immobilized partner concentration (10-6M): 132
P value: 0.75
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: PLEKHA7
Fragment: 626-635
Site: 14-3-3_pSer631
Modification: (SEP)631
Sequence: HVDRRSMPSM
Construct: HVDRR(pSer)MPSM (crude synthetic peptide)

Protein B

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta
Average holdup BI: 0.24
Holdup BI standard deviation: 0.11
Immobilized partner concentration (10-6M): 132
P value: 2.24
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.37
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: PLEKHA7
Fragment: 981-990
Site: 14-3-3_pSer986
Modification: (SEP)986
Sequence: LRSYVSEPEL
Construct: LRSYV(pSer)EPEL (crude synthetic peptide)

Protein B

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta
Average holdup BI: 0.59
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 132
P value: 4.42
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.04
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: PLEKHA7
Fragment: 1008-1017
Site: 14-3-3_pThr1013
Modification: (TPO)1013
Sequence: ESRYQTLPGR
Construct: ESRYQ(pThr)LPGR (crude synthetic peptide)

Protein B

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta
Average holdup BI: 0.66
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 132
P value: 6.66
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.17
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: 1032-1041
Site: 14-3-3_pThr1037
Modification: (TPO)1037
Sequence: IAPYVTLRRG
Construct: IAPYV(pThr)LRRG (crude synthetic peptide)

Protein B

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta
Average holdup BI: 0.28
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 132
P value: 4.47
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.46
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: PLEKHA7
Fragment: 467-476
Site: 14-3-3_pThr472
Modification: (TPO)472
Sequence: PENYQTLPKS
Construct: PENYQ(pThr)LPKS (crude synthetic peptide)

Protein B

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta
Average holdup BI: 0.2
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 132
P value: 3.87
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: 533-542
Site: 14-3-3_pThr538
Modification: (TPO)538
Sequence: RHGSPTAPI-
Construct: FRHGSP(pThr)API (crude synthetic peptide)

Protein B

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta
Measured dissociation constant (10-6M): <10
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: 5
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: 897-906
Site: 14-3-3_pThr902
Modification: (TPO)902
Sequence: QLRKVTSPLQ
Construct: QLRKV(pThr)SPLQ (crude synthetic peptide)

Protein B

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta
Average holdup BI: 0.14
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 132
P value: 2.59
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))