Interaction Details


Details of experiments between P31947 and Q5TCZ1
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: SH3PXD2A
Fragment: 997-1006
Site: 14-3-3_pSer1002
Modification: (SEP)1002
Sequence: LRRNESLTAT
Construct: LRRNE(pSer)LTAT (crude synthetic peptide)
Average holdup BI: -0.14
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 113
P value: 2.51
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 2

Protein A

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

Protein B

Protein: SH3PXD2A
Fragment: 997-1006
Site: 14-3-3_pSer1002
Modification: (SEP)1002
Sequence: LRRNESLTAT
Construct: LRRNE(pSer)LTAT (crude synthetic peptide)
Average holdup BI: 0.01
Holdup BI standard deviation: 0.11
Immobilized partner concentration (10-6M): 142
P value: 0.06
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: SH3PXD2A
Fragment: 997-1006
Site: 14-3-3_pSer1002
Modification: (SEP)1002
Sequence: LRRNESLTAT
Construct: LRRNE(pSer)LTAT (crude synthetic peptide)
Average holdup BI: -0.06
Holdup BI standard deviation: 0.14
Immobilized partner concentration (10-6M): 111
P value: 0.39
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))

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: SH3PXD2A
Fragment: 1012-1021
Site: 14-3-3_pSer1017
Modification: (SEP)1017
Sequence: VRRNSSFSTA
Construct: VRRNS(pSer)FSTA (crude synthetic peptide)
Average holdup BI: 0.01
Holdup BI standard deviation: 0.2
Immobilized partner concentration (10-6M): 113
P value: 0.02
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: SH3PXD2A
Fragment: 1012-1021
Site: 14-3-3_pSer1017
Modification: (SEP)1017
Sequence: VRRNSSFSTA
Construct: VRRNS(pSer)FSTA (crude synthetic peptide)
Average holdup BI: 0.06
Holdup BI standard deviation: 0.25
Immobilized partner concentration (10-6M): 142
P value: 0.19
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: SH3PXD2A
Fragment: 1012-1021
Site: 14-3-3_pSer1017
Modification: (SEP)1017
Sequence: VRRNSSFSTA
Construct: VRRNS(pSer)FSTA (crude synthetic peptide)
Average holdup BI: -0.32
Holdup BI standard deviation: 0.87
Immobilized partner concentration (10-6M): 111
P value: 0.41
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: SH3PXD2A
Fragment: 1047-1056
Site: 14-3-3_pSer1052
Modification: (SEP)1052
Sequence: PAQRNSIPVS
Construct: PAQRN(pSer)IPVS (crude synthetic peptide)
Average holdup BI: 0.02
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 113
P value: 1.48
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: SH3PXD2A
Fragment: 1047-1056
Site: 14-3-3_pSer1052
Modification: (SEP)1052
Sequence: PAQRNSIPVS
Construct: PAQRN(pSer)IPVS (crude synthetic peptide)
Average holdup BI: 0
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 142
P value: 0.15
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: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: SH3PXD2A
Fragment: 1047-1056
Site: 14-3-3_pSer1052
Modification: (SEP)1052
Sequence: PAQRNSIPVS
Construct: PAQRN(pSer)IPVS (crude synthetic peptide)
Average holdup BI: -0.02
Holdup BI standard deviation: 0.15
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 10

Protein A

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

Protein B

Protein: SH3PXD2A
Fragment: 374-383
Site: 14-3-3_pSer379
Modification: (SEP)379
Sequence: AKKEISLPIL
Construct: AKKEI(pSer)LPIL (crude synthetic peptide)
Average holdup BI: 0.31
Holdup BI standard deviation: 0.16
Immobilized partner concentration (10-6M): 113
P value: 1.92
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 11

Protein A

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

Protein B

Protein: SH3PXD2A
Fragment: 374-383
Site: 14-3-3_pSer379
Modification: (SEP)379
Sequence: AKKEISLPIL
Construct: AKKEI(pSer)LPIL (crude synthetic peptide)
Average holdup BI: 0.32
Holdup BI standard deviation: 0.16
Immobilized partner concentration (10-6M): 142
P value: 2.02
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.52
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: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: SH3PXD2A
Fragment: 374-383
Site: 14-3-3_pSer379
Modification: (SEP)379
Sequence: AKKEISLPIL
Construct: AKKEI(pSer)LPIL (crude synthetic peptide)
Average holdup BI: 0.32
Holdup BI standard deviation: 0.09
Immobilized partner concentration (10-6M): 111
P value: 2.98
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.63
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: SH3PXD2A
Fragment: 415-424
Site: 14-3-3_pSer420
Modification: (SEP)420
Sequence: QRAQISSPNL
Construct: QRAQI(pSer)SPNL (crude synthetic peptide)
Average holdup BI: 0.34
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 142
P value: 4.26
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.55
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: SH3PXD2A
Fragment: 415-424
Site: 14-3-3_pSer420
Modification: (SEP)420
Sequence: QRAQISSPNL
Construct: QRAQI(pSer)SPNL (crude synthetic peptide)
Average holdup BI: 0.26
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 113
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: 3.5
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
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: SH3PXD2A
Fragment: 415-424
Site: 14-3-3_pSer420
Modification: (SEP)420
Sequence: QRAQISSPNL
Construct: QRAQI(pSer)SPNL (crude synthetic peptide)
Average holdup BI: 0.19
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 111
P value: 3.49
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 16

Protein A

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

Protein B

Protein: SH3PXD2A
Fragment: 562-571
Site: 14-3-3_pSer567
Modification: (SEP)567
Sequence: AFGFDSEPEL
Construct: AFGFD(pSer)EPEL (crude synthetic peptide)
Average holdup BI: 0.11
Holdup BI standard deviation: 0.41
Immobilized partner concentration (10-6M): 113
P value: 0.21
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: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: SH3PXD2A
Fragment: 562-571
Site: 14-3-3_pSer567
Modification: (SEP)567
Sequence: AFGFDSEPEL
Construct: AFGFD(pSer)EPEL (crude synthetic peptide)
Average holdup BI: -0.05
Holdup BI standard deviation: 0.45
Immobilized partner concentration (10-6M): 111
P value: 0.08
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 18

Protein A

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

Protein B

Protein: SH3PXD2A
Fragment: 633-642
Site: 14-3-3_pSer638
Modification: (SEP)638
Sequence: LSARGSSGDS
Construct: LSARG(pSer)SGDS (crude synthetic peptide)
Average holdup BI: 0.11
Holdup BI standard deviation: 0.33
Immobilized partner concentration (10-6M): 113
P value: 0.27
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 19

Protein A

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

Protein B

Protein: SH3PXD2A
Fragment: 633-642
Site: 14-3-3_pSer638
Modification: (SEP)638
Sequence: LSARGSSGDS
Construct: LSARG(pSer)SGDS (crude synthetic peptide)
Average holdup BI: 0.18
Holdup BI standard deviation: 0.41
Immobilized partner concentration (10-6M): 142
P value: 0.37
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 20

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: SH3PXD2A
Fragment: 633-642
Site: 14-3-3_pSer638
Modification: (SEP)638
Sequence: LSARGSSGDS
Construct: LSARG(pSer)SGDS (crude synthetic peptide)
Average holdup BI: 0.18
Holdup BI standard deviation: 1.1
Immobilized partner concentration (10-6M): 111
P value: 0.11
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: SH3PXD2A
Fragment: 251-260
Site: 14-3-3_pThr256
Modification: (TPO)256
Sequence: LDRRWTLGGM
Construct: LDRRW(pThr)LGGM (crude synthetic peptide)
Average holdup BI: 0.68
Holdup BI standard deviation: 0.1
Immobilized partner concentration (10-6M): 142
P value: 3.7
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 22

Protein A

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

Protein B

Protein: SH3PXD2A
Fragment: 251-260
Site: 14-3-3_pThr256
Modification: (TPO)256
Sequence: LDRRWTLGGM
Construct: LDRRW(pThr)LGGM (crude synthetic peptide)
Measured dissociation constant (10-6M): <100
Immobilized partner concentration (10-6M): 113
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
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 4))

Experiment series 23

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: SH3PXD2A
Fragment: 251-260
Site: 14-3-3_pThr256
Modification: (TPO)256
Sequence: LDRRWTLGGM
Construct: LDRRW(pThr)LGGM (crude synthetic peptide)
Average holdup BI: 0.79
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 111
P value: 1.97
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.53
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: SH3PXD2A
Fragment: 513-522
Site: 14-3-3_pThr518
Modification: (TPO)518
Sequence: RTSTLTRPKV
Construct: RTSTL(pThr)RPKV (crude synthetic peptide)
Average holdup BI: -0.03
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 113
P value: 0.57
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 25

Protein A

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

Protein B

Protein: SH3PXD2A
Fragment: 513-522
Site: 14-3-3_pThr518
Modification: (TPO)518
Sequence: RTSTLTRPKV
Construct: RTSTL(pThr)RPKV (crude synthetic peptide)
Average holdup BI: 0.04
Holdup BI standard deviation: 0.05
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 26

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: SH3PXD2A
Fragment: 513-522
Site: 14-3-3_pThr518
Modification: (TPO)518
Sequence: RTSTLTRPKV
Construct: RTSTL(pThr)RPKV (crude synthetic peptide)
Average holdup BI: 0.07
Holdup BI standard deviation: 0.09
Immobilized partner concentration (10-6M): 111
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))