Interaction Details


Details of experiments between Q9H0B6 and P62258
Note that the results of all experiments are listed, regardless of the modification states of the fragments.


Experiment series 1

Protein A

Protein: KLC2
Fragment: 540-549
Site: 14-3-3_pSer545
Modification: (SEP)545
Sequence: LRRSGSFGKL
Construct: LRRSG(pSer)FGKL (crude synthetic peptide)

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.02
Holdup BI standard deviation: 0.12
Immobilized partner concentration (10-6M): 116
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 2

Protein A

Protein: KLC2
Fragment: 540-549
Site: 14-3-3_pSer545
Modification: (SEP)545
Sequence: LRRSGSFGKL
Construct: LRRSG(pSer)FGKL (crude synthetic peptide)

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.09
Holdup BI standard deviation: 0.11
Immobilized partner concentration (10-6M): 143
P value: 0.8
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: KLC2
Fragment: 577-586
Site: 14-3-3_pSer582
Modification: (SEP)582
Sequence: MKRASSLNFL
Construct: MKRAS(pSer)LNFL (crude synthetic peptide)

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.74
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 143
P value: 5.89
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.3
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 4

Protein A

Protein: KLC2
Fragment: 577-586
Site: 14-3-3_pSer582
Modification: (SEP)582
Sequence: MKRASSLNFL
Construct: MKRAS(pSer)LNFL (crude synthetic peptide)

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.62
Holdup BI standard deviation: 0.1
Immobilized partner concentration (10-6M): 116
P value: 2.87
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.15
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: KLC2
Fragment: 584-593
Site: 14-3-3_pSer589
Modification: (SEP)589
Sequence: NFLNKSVEEP
Construct: NFLNK(pSer)VEEP (crude synthetic peptide)

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.58
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 143
P value: 3.09
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.98
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: KLC2
Fragment: 584-593
Site: 14-3-3_pSer589
Modification: (SEP)589
Sequence: NFLNKSVEEP
Construct: NFLNK(pSer)VEEP (crude synthetic peptide)

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.23
Holdup BI standard deviation: 0.37
Immobilized partner concentration (10-6M): 116
P value: 0.46
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: KLC2
Fragment: 603-612
Site: 14-3-3_pSer608
Modification: (SEP)608
Sequence: DSRTLSSSSM
Construct: DSRTL(pSer)SSSM (crude synthetic peptide)

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.24
Holdup BI standard deviation: 0.25
Immobilized partner concentration (10-6M): 116
P value: 0.93
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: KLC2
Fragment: 603-612
Site: 14-3-3_pSer608
Modification: (SEP)608
Sequence: DSRTLSSSSM
Construct: DSRTL(pSer)SSSM (crude synthetic peptide)

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.07
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 143
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 9

Protein A

Protein: KLC2
Fragment: 605-614
Site: 14-3-3_pSer610
Modification: (SEP)610
Sequence: RTLSSSSMDL
Construct: RTLSS(pSer)SMDL (crude synthetic peptide)

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.19
Holdup BI standard deviation: 0.17
Immobilized partner concentration (10-6M): 116
P value: 1.09
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: KLC2
Fragment: 605-614
Site: 14-3-3_pSer610
Modification: (SEP)610
Sequence: RTLSSSSMDL
Construct: RTLSS(pSer)SMDL (crude synthetic peptide)

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.09
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 143
P value: 1.36
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 11

Protein A

Protein: KLC2
Fragment: 610-619
Site: 14-3-3_pSer615
Modification: (SEP)615
Sequence: SSMDLSRRSS
Construct: SSMDL(pSer)RRSS (crude synthetic peptide)

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.11
Holdup BI standard deviation: 0.19
Immobilized partner concentration (10-6M): 116
P value: 0.54
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: KLC2
Fragment: 610-619
Site: 14-3-3_pSer615
Modification: (SEP)615
Sequence: SSMDLSRRSS
Construct: SSMDL(pSer)RRSS (crude synthetic peptide)

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.01
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 143
P value: 0.13
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: KLC2
Fragment: 614-623
Site: 14-3-3_pSer619
Modification: (SEP)619
Sequence: LSRRSSLVG-
Construct: DLSRRS(pSer)LVG (crude synthetic peptide)

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.05
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 116
P value: 1.89
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: KLC2
Fragment: 614-623
Site: 14-3-3_pSer619
Modification: (SEP)619
Sequence: LSRRSSLVG-
Construct: DLSRRS(pSer)LVG (crude synthetic peptide)

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.1
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 143
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))