Interaction Details


Details of experiments between P31947 and Q9UJF2
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: RASAL2
Fragment: 129-138
Site: 14-3-3_pSer134
Modification: (SEP)134
Sequence: LRRTVSVPSE
Construct: LRRTV(pSer)VPSE (crude synthetic peptide)
Average holdup BI: 0.51
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 142
P value: 5.83
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.86
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: RASAL2
Fragment: 129-138
Site: 14-3-3_pSer134
Modification: (SEP)134
Sequence: LRRTVSVPSE
Construct: LRRTV(pSer)VPSE (crude synthetic peptide)
Average holdup BI: 0.44
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 113
P value: 6.34
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.83
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

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: RASAL2
Fragment: 129-138
Site: 14-3-3_pSer134
Modification: (SEP)134
Sequence: LRRTVSVPSE
Construct: LRRTV(pSer)VPSE (crude synthetic peptide)
Average holdup BI: 0.59
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 111
P value: 6.32
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.12
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: RASAL2
Fragment: 695-704
Site: 14-3-3_pSer700
Modification: (SEP)700
Sequence: DERESSLPNG
Construct: DERES(pSer)LPNG (crude synthetic peptide)
Average holdup BI: -0.11
Holdup BI standard deviation: 0.17
Immobilized partner concentration (10-6M): 113
P value: 0.69
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
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: RASAL2
Fragment: 695-704
Site: 14-3-3_pSer700
Modification: (SEP)700
Sequence: DERESSLPNG
Construct: DERES(pSer)LPNG (crude synthetic peptide)
Average holdup BI: -0.03
Holdup BI standard deviation: 0.08
Immobilized partner concentration (10-6M): 111
P value: 0.36
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: RASAL2
Fragment: 731-740
Site: 14-3-3_pSer736
Modification: (SEP)736
Sequence: IRLTGSQLSI
Construct: IRLTG(pSer)QLSI (crude synthetic peptide)
Average holdup BI: -0.02
Holdup BI standard deviation: 0.24
Immobilized partner concentration (10-6M): 113
P value: 0.05
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
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: RASAL2
Fragment: 731-740
Site: 14-3-3_pSer736
Modification: (SEP)736
Sequence: IRLTGSQLSI
Construct: IRLTG(pSer)QLSI (crude synthetic peptide)
Average holdup BI: 0.01
Holdup BI standard deviation: 0.1
Immobilized partner concentration (10-6M): 111
P value: 0.04
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: RASAL2
Fragment: 734-743
Site: 14-3-3_pSer739
Modification: (SEP)739
Sequence: TGSQLSITQV
Construct: TGSQL(pSer)ITQV (crude synthetic peptide)
Average holdup BI: 0.04
Holdup BI standard deviation: 0.08
Immobilized partner concentration (10-6M): 113
P value: 0.45
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
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: RASAL2
Fragment: 734-743
Site: 14-3-3_pSer739
Modification: (SEP)739
Sequence: TGSQLSITQV
Construct: TGSQL(pSer)ITQV (crude synthetic peptide)
Average holdup BI: 0.05
Holdup BI standard deviation: 0.11
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 10

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: RASAL2
Fragment: 734-743
Site: 14-3-3_pSer739
Modification: (SEP)739
Sequence: TGSQLSITQV
Construct: TGSQL(pSer)ITQV (crude synthetic peptide)
Average holdup BI: 0.04
Holdup BI standard deviation: 0.09
Immobilized partner concentration (10-6M): 111
P value: 0.35
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 11

Protein A

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

Protein B

Protein: RASAL2
Fragment: 836-845
Site: 14-3-3_pSer841
Modification: (SEP)841
Sequence: KRSTQSEDFS
Construct: KRSTQ(pSer)EDFS (crude synthetic peptide)
Average holdup BI: -0.11
Holdup BI standard deviation: 0.31
Immobilized partner concentration (10-6M): 113
P value: 0.18
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))

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: RASAL2
Fragment: 836-845
Site: 14-3-3_pSer841
Modification: (SEP)841
Sequence: KRSTQSEDFS
Construct: KRSTQ(pSer)EDFS (crude synthetic peptide)
Average holdup BI: 0.07
Holdup BI standard deviation: 0.3
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 13

Protein A

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

Protein B

Protein: RASAL2
Fragment: 882-891
Site: 14-3-3_pSer887
Modification: (SEP)887
Sequence: AKAPPSLPHS
Construct: AKAPP(pSer)LPHS (crude synthetic peptide)
Average holdup BI: 0.07
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 113
P value: 0.99
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: RASAL2
Fragment: 882-891
Site: 14-3-3_pSer887
Modification: (SEP)887
Sequence: AKAPPSLPHS
Construct: AKAPP(pSer)LPHS (crude synthetic peptide)
Average holdup BI: -0.06
Holdup BI standard deviation: 0.2
Immobilized partner concentration (10-6M): 142
P value: 0.29
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: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: RASAL2
Fragment: 882-891
Site: 14-3-3_pSer887
Modification: (SEP)887
Sequence: AKAPPSLPHS
Construct: AKAPP(pSer)LPHS (crude synthetic peptide)
Average holdup BI: -0.04
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 111
P value: 0.71
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: RASAL2
Fragment: 84-93
Site: 14-3-3_pSer89
Modification: (SEP)89
Sequence: LKESRSHESL
Construct: LKESR(pSer)HESL (crude synthetic peptide)
Average holdup BI: 0.11
Holdup BI standard deviation: 0.16
Immobilized partner concentration (10-6M): 113
P value: 0.56
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: RASAL2
Fragment: 84-93
Site: 14-3-3_pSer89
Modification: (SEP)89
Sequence: LKESRSHESL
Construct: LKESR(pSer)HESL (crude synthetic peptide)
Average holdup BI: 0.07
Holdup BI standard deviation: 0.19
Immobilized partner concentration (10-6M): 142
P value: 0.31
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))

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: RASAL2
Fragment: 84-93
Site: 14-3-3_pSer89
Modification: (SEP)89
Sequence: LKESRSHESL
Construct: LKESR(pSer)HESL (crude synthetic peptide)
Average holdup BI: 0.01
Holdup BI standard deviation: 0.18
Immobilized partner concentration (10-6M): 111
P value: 0.04
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: RASAL2
Fragment: 612-621
Site: 14-3-3_pThr617
Modification: (TPO)617
Sequence: ITKSLTNPTP
Construct: ITKSL(pThr)NPTP (crude synthetic peptide)
Average holdup BI: 0.51
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 113
P value: 6.94
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 20

Protein A

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

Protein B

Protein: RASAL2
Fragment: 612-621
Site: 14-3-3_pThr617
Modification: (TPO)617
Sequence: ITKSLTNPTP
Construct: ITKSL(pThr)NPTP (crude synthetic peptide)
Average holdup BI: 0.57
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 142
P value: 6.63
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.97
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: RASAL2
Fragment: 612-621
Site: 14-3-3_pThr617
Modification: (TPO)617
Sequence: ITKSLTNPTP
Construct: ITKSL(pThr)NPTP (crude synthetic peptide)
Average holdup BI: 0.5
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 111
P value: 7.3
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.96
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

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: RASAL2
Fragment: 844-853
Site: 14-3-3_pThr849
Modification: (TPO)849
Sequence: FSRRHTVPDR
Construct: FSRRH(pThr)VPDR (crude synthetic peptide)
Average holdup BI: 0.71
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 113
P value: 6.33
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.34
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: RASAL2
Fragment: 844-853
Site: 14-3-3_pThr849
Modification: (TPO)849
Sequence: FSRRHTVPDR
Construct: FSRRH(pThr)VPDR (crude synthetic peptide)
Average holdup BI: 0.72
Holdup BI standard deviation: 0.1
Immobilized partner concentration (10-6M): 142
P value: 3.27
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.26
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: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: RASAL2
Fragment: 844-853
Site: 14-3-3_pThr849
Modification: (TPO)849
Sequence: FSRRHTVPDR
Construct: FSRRH(pThr)VPDR (crude synthetic peptide)
Average holdup BI: 0.69
Holdup BI standard deviation: 0.08
Immobilized partner concentration (10-6M): 111
P value: 3.2
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.3
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: RASAL2
Fragment: 936-945
Site: 14-3-3_pThr941
Modification: (TPO)941
Sequence: IQRQQTQQVQ
Construct: IQRQQ(pThr)QQVQ (crude synthetic peptide)
Average holdup BI: 0.05
Holdup BI standard deviation: 0.21
Immobilized partner concentration (10-6M): 113
P value: 0.17
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: RASAL2
Fragment: 936-945
Site: 14-3-3_pThr941
Modification: (TPO)941
Sequence: IQRQQTQQVQ
Construct: IQRQQ(pThr)QQVQ (crude synthetic peptide)
Average holdup BI: 0.03
Holdup BI standard deviation: 0.18
Immobilized partner concentration (10-6M): 142
P value: 0.11
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 27

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: RASAL2
Fragment: 936-945
Site: 14-3-3_pThr941
Modification: (TPO)941
Sequence: IQRQQTQQVQ
Construct: IQRQQ(pThr)QQVQ (crude synthetic peptide)
Average holdup BI: -0.21
Holdup BI standard deviation: 0.4
Immobilized partner concentration (10-6M): 111
P value: 0.56
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))