Interaction Details


Details of experiments between P31947 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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

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
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 113
P value: 0.09
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

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
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 142
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: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: -0.01
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 111
P value: 0.09
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: 0.05
Holdup BI standard deviation: 0.19
Immobilized partner concentration (10-6M): 113
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 5

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: 0.1
Holdup BI standard deviation: 0.1
Immobilized partner concentration (10-6M): 142
P value: 0.94
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 6

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: 0.12
Holdup BI standard deviation: 0.13
Immobilized partner concentration (10-6M): 111
P value: 0.9
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 7

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: 0.92
Holdup BI standard deviation: 0.09
Immobilized partner concentration (10-6M): 142
P value: 2.34
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.89
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: 0.19
Holdup BI standard deviation: 0.36
Immobilized partner concentration (10-6M): 113
P value: 0.25
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 9

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: -0.19
Holdup BI standard deviation: 0.53
Immobilized partner concentration (10-6M): 111
P value: 0.26
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 10

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: 0.77
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 142
P value: 8.28
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 11

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: 0.72
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 113
P value: 9.69
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.35
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

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): 88
Standard deviation of dissociation constant (10-6M): 7
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: 4.06
DOI: 10.1101/2022.12.08.519103

Experiment series 13

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: 0.67
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 111
P value: 7.98
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 14

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: 0.74
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 113
P value: 4.06
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.4
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: 0.77
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 142
P value: 7.26
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.36
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: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: 0.97
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 111
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: 5.42
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: 0.01
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 113
P value: 0.09
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 18

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: 0.03
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 142
P value: 0.86
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
The affinity was below the detection threshold of the assay.
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 19

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: 0.04
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 111
P value: 0.95
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 20

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: -0.04
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 113
P value: 0.55
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 21

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: -0.11
Holdup BI standard deviation: 0.09
Immobilized partner concentration (10-6M): 142
P value: 1.54
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 22

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: 0.04
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 111
P value: 0.95
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: PLEKHA7
Fragment: 626-635
Site: 14-3-3_pSer631
Modification: (SEP)631
Sequence: HVDRRSMPSM
Construct: HVDRR(pSer)MPSM (crude synthetic peptide)
Average holdup BI: 0.2
Holdup BI standard deviation: 0.7
Immobilized partner concentration (10-6M): 113
P value: 0.14
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: PLEKHA7
Fragment: 626-635
Site: 14-3-3_pSer631
Modification: (SEP)631
Sequence: HVDRRSMPSM
Construct: HVDRR(pSer)MPSM (crude synthetic peptide)
Average holdup BI: 0.05
Holdup BI standard deviation: 0.09
Immobilized partner concentration (10-6M): 142
P value: 0.59
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))

Experiment series 25

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: PLEKHA7
Fragment: 626-635
Site: 14-3-3_pSer631
Modification: (SEP)631
Sequence: HVDRRSMPSM
Construct: HVDRR(pSer)MPSM (crude synthetic peptide)
Average holdup BI: -0.27
Holdup BI standard deviation: 1.46
Immobilized partner concentration (10-6M): 111
P value: 0.15
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: 0.08
Holdup BI standard deviation: 0.12
Immobilized partner concentration (10-6M): 113
P value: 0.63
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: 0.09
Holdup BI standard deviation: 0.14
Immobilized partner concentration (10-6M): 142
P value: 0.64
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 28

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: 0.16
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 111
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
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 29

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: 0.59
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 113
P value: 7.12
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.11
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: 0.63
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 142
P value: 7.03
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.09
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: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: 0.59
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 111
P value: 5.82
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.11
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: 0.11
Holdup BI standard deviation: 0.09
Immobilized partner concentration (10-6M): 113
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 33

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: 0.13
Holdup BI standard deviation: 0.09
Immobilized partner concentration (10-6M): 142
P value: 1.69
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: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: 0.12
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 111
P value: 1.95
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: 0.13
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 113
P value: 3.66
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: 0.15
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 142
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
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 37

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: 0.19
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 111
P value: 4.44
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 38

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

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): 142
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 39

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: 0.78
Holdup BI standard deviation: 0.15
Immobilized partner concentration (10-6M): 113
P value: 4.03
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.48
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: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

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): 111
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
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 4))

Experiment series 41

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: 0.08
Holdup BI standard deviation: 0.25
Immobilized partner concentration (10-6M): 113
P value: 0.26
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: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: -0.03
Holdup BI standard deviation: 0.14
Immobilized partner concentration (10-6M): 111
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 43

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: 0.04
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 113
P value: 0.5
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: 0.06
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 142
P value: 1.3
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: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

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: 0.07
Immobilized partner concentration (10-6M): 111
P value: 1.18
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))