Interaction Details


Details of experiments between Q53ET0 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: CRTC2
Fragment: 122-131
Site: 14-3-3_pSer127
Modification: (SEP)127
Sequence: TRHIDSSPYS
Construct: TRHID(pSer)SPYS (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: 0.29
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 143
P value: 4.7
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.46
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: CRTC2
Fragment: 122-131
Site: 14-3-3_pSer127
Modification: (SEP)127
Sequence: TRHIDSSPYS
Construct: TRHID(pSer)SPYS (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: 0.18
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 116
P value: 2.81
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: CRTC2
Fragment: 166-175
Site: 14-3-3_pSer171
Modification: (SEP)171
Sequence: LNRTSSDSAL
Construct: LNRTS(pSer)DSAL (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: 0.22
Holdup BI standard deviation: 0.09
Immobilized partner concentration (10-6M): 143
P value: 2.62
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.31
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: CRTC2
Fragment: 166-175
Site: 14-3-3_pSer171
Modification: (SEP)171
Sequence: LNRTSSDSAL
Construct: LNRTS(pSer)DSAL (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: 0.22
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 116
P value: 6.04
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 5

Protein A

Protein: CRTC2
Fragment: 442-451
Site: 14-3-3_pSer447
Modification: (SEP)447
Sequence: ADARRSQQQL
Construct: ADARR(pSer)QQQL (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: 0.25
Holdup BI standard deviation: 0.34
Immobilized partner concentration (10-6M): 116
P value: 0.44
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: CRTC2
Fragment: 442-451
Site: 14-3-3_pSer447
Modification: (SEP)447
Sequence: ADARRSQQQL
Construct: ADARR(pSer)QQQL (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: 0.06
Holdup BI standard deviation: 1.36
Immobilized partner concentration (10-6M): 143
P value: 0.03
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 7

Protein A

Protein: CRTC2
Fragment: 487-496
Site: 14-3-3_pSer492
Modification: (SEP)492
Sequence: PYSSPSLVLP
Construct: PYSSP(pSer)LVLP (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: -0.12
Holdup BI standard deviation: 0.17
Immobilized partner concentration (10-6M): 116
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 8

Protein A

Protein: CRTC2
Fragment: 487-496
Site: 14-3-3_pSer492
Modification: (SEP)492
Sequence: PYSSPSLVLP
Construct: PYSSP(pSer)LVLP (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: -0.21
Holdup BI standard deviation: 0.4
Immobilized partner concentration (10-6M): 143
P value: 0.62
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: CRTC2
Fragment: 65-74
Site: 14-3-3_pSer70
Modification: (SEP)70
Sequence: SHYGGSLPNV
Construct: SHYGG(pSer)LPNV (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: 0.42
Holdup BI standard deviation: 0.17
Immobilized partner concentration (10-6M): 116
P value: 2.4
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.8
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: CRTC2
Fragment: 65-74
Site: 14-3-3_pSer70
Modification: (SEP)70
Sequence: SHYGGSLPNV
Construct: SHYGG(pSer)LPNV (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: 0.47
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 143
P value: 6
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.8
DOI: 10.1101/2022.12.08.519103
Experimental Details

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