Interaction Details


Details of experiments between P31947 and Q7Z401
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: DENND4A
Fragment: 1030-1039
Site: 14-3-3_pSer1035
Modification: (SEP)1035
Sequence: RNRNLSGGVL
Construct: RNRNL(pSer)GGVL (crude synthetic peptide)
Average holdup BI: 0.05
Holdup BI standard deviation: 0.09
Immobilized partner concentration (10-6M): 113
P value: 0.52
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: DENND4A
Fragment: 1030-1039
Site: 14-3-3_pSer1035
Modification: (SEP)1035
Sequence: RNRNLSGGVL
Construct: RNRNL(pSer)GGVL (crude synthetic peptide)
Average holdup BI: 0.11
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 142
P value: 1.84
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: DENND4A
Fragment: 1030-1039
Site: 14-3-3_pSer1035
Modification: (SEP)1035
Sequence: RNRNLSGGVL
Construct: RNRNL(pSer)GGVL (crude synthetic peptide)
Average holdup BI: 0.01
Holdup BI standard deviation: 0.68
Immobilized partner concentration (10-6M): 111
P value: 0.01
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: DENND4A
Fragment: 1146-1155
Site: 14-3-3_pSer1151
Modification: (SEP)1151
Sequence: IQRMNSSFSV
Construct: IQRMN(pSer)SFSV (crude synthetic peptide)
Average holdup BI: 0.16
Holdup BI standard deviation: 0.16
Immobilized partner concentration (10-6M): 113
P value: 0.97
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: DENND4A
Fragment: 1146-1155
Site: 14-3-3_pSer1151
Modification: (SEP)1151
Sequence: IQRMNSSFSV
Construct: IQRMN(pSer)SFSV (crude synthetic peptide)
Average holdup BI: 0.04
Holdup BI standard deviation: 0.09
Immobilized partner concentration (10-6M): 142
P value: 0.44
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: DENND4A
Fragment: 1146-1155
Site: 14-3-3_pSer1151
Modification: (SEP)1151
Sequence: IQRMNSSFSV
Construct: IQRMN(pSer)SFSV (crude synthetic peptide)
Average holdup BI: 0.05
Holdup BI standard deviation: 0.26
Immobilized partner concentration (10-6M): 111
P value: 0.13
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: DENND4A
Fragment: 1220-1229
Site: 14-3-3_pSer1225
Modification: (SEP)1225
Sequence: TSRTPSIDLQ
Construct: TSRTP(pSer)IDLQ (crude synthetic peptide)
Average holdup BI: 0.03
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 113
P value: 1.21
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 8

Protein A

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

Protein B

Protein: DENND4A
Fragment: 1220-1229
Site: 14-3-3_pSer1225
Modification: (SEP)1225
Sequence: TSRTPSIDLQ
Construct: TSRTP(pSer)IDLQ (crude synthetic peptide)
Average holdup BI: 0.02
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 142
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 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: DENND4A
Fragment: 1220-1229
Site: 14-3-3_pSer1225
Modification: (SEP)1225
Sequence: TSRTPSIDLQ
Construct: TSRTP(pSer)IDLQ (crude synthetic peptide)
Average holdup BI: -0.04
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 111
P value: 1.2
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: DENND4A
Fragment: 1276-1285
Site: 14-3-3_pSer1281
Modification: (SEP)1281
Sequence: RDRLWSSPAF
Construct: RDRLW(pSer)SPAF (crude synthetic peptide)
Average holdup BI: 0.84
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 142
P value: 3.53
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.56
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: DENND4A
Fragment: 1276-1285
Site: 14-3-3_pSer1281
Modification: (SEP)1281
Sequence: RDRLWSSPAF
Construct: RDRLW(pSer)SPAF (crude synthetic peptide)
Average holdup BI: 0.53
Holdup BI standard deviation: 0.18
Immobilized partner concentration (10-6M): 113
P value: 1.72
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.99
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
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: DENND4A
Fragment: 1276-1285
Site: 14-3-3_pSer1281
Modification: (SEP)1281
Sequence: RDRLWSSPAF
Construct: RDRLW(pSer)SPAF (crude synthetic peptide)
Average holdup BI: 0.67
Holdup BI standard deviation: 0.11
Immobilized partner concentration (10-6M): 111
P value: 2.66
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 13

Protein A

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

Protein B

Protein: DENND4A
Fragment: 1503-1512
Site: 14-3-3_pSer1508
Modification: (SEP)1508
Sequence: RYFLKSSPST
Construct: RYFLK(pSer)SPST (crude synthetic peptide)
Average holdup BI: 0.63
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 142
P value: 6.44
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.08
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: DENND4A
Fragment: 1503-1512
Site: 14-3-3_pSer1508
Modification: (SEP)1508
Sequence: RYFLKSSPST
Construct: RYFLK(pSer)SPST (crude synthetic peptide)
Average holdup BI: 0.53
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 113
P value: 5.74
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.01
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: DENND4A
Fragment: 1503-1512
Site: 14-3-3_pSer1508
Modification: (SEP)1508
Sequence: RYFLKSSPST
Construct: RYFLK(pSer)SPST (crude synthetic peptide)
Average holdup BI: 0.51
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 111
P value: 5.77
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.97
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: DENND4A
Fragment: 1586-1595
Site: 14-3-3_pSer1591
Modification: (SEP)1591
Sequence: RSISTSGPLD
Construct: RSIST(pSer)GPLD (crude synthetic peptide)
Average holdup BI: 0.08
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 113
P value: 3.85
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
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: DENND4A
Fragment: 1586-1595
Site: 14-3-3_pSer1591
Modification: (SEP)1591
Sequence: RSISTSGPLD
Construct: RSIST(pSer)GPLD (crude synthetic peptide)
Average holdup BI: 0.12
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 142
P value: 5.65
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 18

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: DENND4A
Fragment: 1586-1595
Site: 14-3-3_pSer1591
Modification: (SEP)1591
Sequence: RSISTSGPLD
Construct: RSIST(pSer)GPLD (crude synthetic peptide)
Average holdup BI: 0.09
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 111
P value: 2.75
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: DENND4A
Fragment: 1585-1594
Site: 14-3-3_pThr1590
Modification: (TPO)1590
Sequence: ARSISTSGPL
Construct: ARSIS(pThr)SGPL (crude synthetic peptide)
Average holdup BI: 0.17
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 113
P value: 5.77
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: DENND4A
Fragment: 1585-1594
Site: 14-3-3_pThr1590
Modification: (TPO)1590
Sequence: ARSISTSGPL
Construct: ARSIS(pThr)SGPL (crude synthetic peptide)
Average holdup BI: 0.22
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 142
P value: 5.07
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 21

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: DENND4A
Fragment: 1585-1594
Site: 14-3-3_pThr1590
Modification: (TPO)1590
Sequence: ARSISTSGPL
Construct: ARSIS(pThr)SGPL (crude synthetic peptide)
Average holdup BI: 0.17
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 111
P value: 4.9
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))