Interaction Details


Details of experiments between P31947 and Q9H0B6
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: KLC2
Fragment: 423-432
Site: 14-3-3_pSer428
Modification: (SEP)428
Sequence: DKRRDSAPYG
Construct: DKRRD(pSer)APYG (crude synthetic peptide)
Average holdup BI: 0.48
Holdup BI standard deviation: 0.21
Immobilized partner concentration (10-6M): 142
P value: 1.45
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.81
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: KLC2
Fragment: 423-432
Site: 14-3-3_pSer428
Modification: (SEP)428
Sequence: DKRRDSAPYG
Construct: DKRRD(pSer)APYG (crude synthetic peptide)
Average holdup BI: -0.23
Holdup BI standard deviation: 1.72
Immobilized partner concentration (10-6M): 113
P value: 0.07
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 3

Protein A

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

Protein B

Protein: KLC2
Fragment: 540-549
Site: 14-3-3_pSer545
Modification: (SEP)545
Sequence: LRRSGSFGKL
Construct: LRRSG(pSer)FGKL (crude synthetic peptide)
Average holdup BI: 0.02
Holdup BI standard deviation: 0.07
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 4

Protein A

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

Protein B

Protein: KLC2
Fragment: 540-549
Site: 14-3-3_pSer545
Modification: (SEP)545
Sequence: LRRSGSFGKL
Construct: LRRSG(pSer)FGKL (crude synthetic peptide)
Average holdup BI: 0.02
Holdup BI standard deviation: 0.14
Immobilized partner concentration (10-6M): 142
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 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: KLC2
Fragment: 540-549
Site: 14-3-3_pSer545
Modification: (SEP)545
Sequence: LRRSGSFGKL
Construct: LRRSG(pSer)FGKL (crude synthetic peptide)
Average holdup BI: 0.1
Holdup BI standard deviation: 0.18
Immobilized partner concentration (10-6M): 111
P value: 0.47
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: KLC2
Fragment: 577-586
Site: 14-3-3_pSer582
Modification: (SEP)582
Sequence: MKRASSLNFL
Construct: MKRAS(pSer)LNFL (crude synthetic peptide)
Average holdup BI: 0.8
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 142
P value: 6.08
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.46
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: KLC2
Fragment: 577-586
Site: 14-3-3_pSer582
Modification: (SEP)582
Sequence: MKRASSLNFL
Construct: MKRAS(pSer)LNFL (crude synthetic peptide)
Average holdup BI: 0.71
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 113
P value: 3.27
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 8

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: KLC2
Fragment: 577-586
Site: 14-3-3_pSer582
Modification: (SEP)582
Sequence: MKRASSLNFL
Construct: MKRAS(pSer)LNFL (crude synthetic peptide)
Average holdup BI: 0.79
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 111
P value: 5.35
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.54
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: KLC2
Fragment: 584-593
Site: 14-3-3_pSer589
Modification: (SEP)589
Sequence: NFLNKSVEEP
Construct: NFLNK(pSer)VEEP (crude synthetic peptide)
Average holdup BI: 0.59
Holdup BI standard deviation: 0.09
Immobilized partner concentration (10-6M): 142
P value: 3.42
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.01
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
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: KLC2
Fragment: 584-593
Site: 14-3-3_pSer589
Modification: (SEP)589
Sequence: NFLNKSVEEP
Construct: NFLNK(pSer)VEEP (crude synthetic peptide)
Average holdup BI: -0.03
Holdup BI standard deviation: 0.15
Immobilized partner concentration (10-6M): 113
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 11

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: KLC2
Fragment: 584-593
Site: 14-3-3_pSer589
Modification: (SEP)589
Sequence: NFLNKSVEEP
Construct: NFLNK(pSer)VEEP (crude synthetic peptide)
Average holdup BI: 0.26
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 111
P value: 2.1
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.49
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: KLC2
Fragment: 603-612
Site: 14-3-3_pSer608
Modification: (SEP)608
Sequence: DSRTLSSSSM
Construct: DSRTL(pSer)SSSM (crude synthetic peptide)
Average holdup BI: 0.25
Holdup BI standard deviation: 0.19
Immobilized partner concentration (10-6M): 113
P value: 1.35
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.48
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: KLC2
Fragment: 603-612
Site: 14-3-3_pSer608
Modification: (SEP)608
Sequence: DSRTLSSSSM
Construct: DSRTL(pSer)SSSM (crude synthetic peptide)
Average holdup BI: 0.08
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 142
P value: 4.03
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: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: KLC2
Fragment: 603-612
Site: 14-3-3_pSer608
Modification: (SEP)608
Sequence: DSRTLSSSSM
Construct: DSRTL(pSer)SSSM (crude synthetic peptide)
Average holdup BI: -0.04
Holdup BI standard deviation: 0.45
Immobilized partner concentration (10-6M): 111
P value: 0.06
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 15

Protein A

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

Protein B

Protein: KLC2
Fragment: 605-614
Site: 14-3-3_pSer610
Modification: (SEP)610
Sequence: RTLSSSSMDL
Construct: RTLSS(pSer)SMDL (crude synthetic peptide)
Average holdup BI: 0.23
Holdup BI standard deviation: 0.12
Immobilized partner concentration (10-6M): 113
P value: 1.9
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.43
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: KLC2
Fragment: 605-614
Site: 14-3-3_pSer610
Modification: (SEP)610
Sequence: RTLSSSSMDL
Construct: RTLSS(pSer)SMDL (crude synthetic peptide)
Average holdup BI: 0.19
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 142
P value: 3.01
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 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: KLC2
Fragment: 605-614
Site: 14-3-3_pSer610
Modification: (SEP)610
Sequence: RTLSSSSMDL
Construct: RTLSS(pSer)SMDL (crude synthetic peptide)
Average holdup BI: 0.08
Holdup BI standard deviation: 0.17
Immobilized partner concentration (10-6M): 111
P value: 0.43
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: KLC2
Fragment: 610-619
Site: 14-3-3_pSer615
Modification: (SEP)615
Sequence: SSMDLSRRSS
Construct: SSMDL(pSer)RRSS (crude synthetic peptide)
Average holdup BI: 0.07
Holdup BI standard deviation: 0.13
Immobilized partner concentration (10-6M): 113
P value: 0.53
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: KLC2
Fragment: 610-619
Site: 14-3-3_pSer615
Modification: (SEP)615
Sequence: SSMDLSRRSS
Construct: SSMDL(pSer)RRSS (crude synthetic peptide)
Average holdup BI: 0
Holdup BI standard deviation: 0.04
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 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: KLC2
Fragment: 610-619
Site: 14-3-3_pSer615
Modification: (SEP)615
Sequence: SSMDLSRRSS
Construct: SSMDL(pSer)RRSS (crude synthetic peptide)
Average holdup BI: -0.08
Holdup BI standard deviation: 0.23
Immobilized partner concentration (10-6M): 111
P value: 0.29
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 21

Protein A

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

Protein B

Protein: KLC2
Fragment: 614-623
Site: 14-3-3_pSer619
Modification: (SEP)619
Sequence: LSRRSSLVG-
Construct: DLSRRS(pSer)LVG (crude synthetic peptide)
Average holdup BI: 0.06
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 113
P value: 1.25
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 22

Protein A

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

Protein B

Protein: KLC2
Fragment: 614-623
Site: 14-3-3_pSer619
Modification: (SEP)619
Sequence: LSRRSSLVG-
Construct: DLSRRS(pSer)LVG (crude synthetic peptide)
Average holdup BI: 0.08
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 142
P value: 1.92
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 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: KLC2
Fragment: 614-623
Site: 14-3-3_pSer619
Modification: (SEP)619
Sequence: LSRRSSLVG-
Construct: DLSRRS(pSer)LVG (crude synthetic peptide)
Average holdup BI: 0.03
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 111
P value: 2.15
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))