Interaction Details


Details of experiments between P31947 and Q53ET0
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: CRTC2
Fragment: 122-131
Site: 14-3-3_pSer127
Modification: (SEP)127
Sequence: TRHIDSSPYS
Construct: TRHID(pSer)SPYS (crude synthetic peptide)
Average holdup BI: 0.13
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 113
P value: 2.18
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 2

Protein A

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

Protein B

Protein: CRTC2
Fragment: 122-131
Site: 14-3-3_pSer127
Modification: (SEP)127
Sequence: TRHIDSSPYS
Construct: TRHID(pSer)SPYS (crude synthetic peptide)
Average holdup BI: 0.19
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 142
P value: 3.65
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 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: CRTC2
Fragment: 122-131
Site: 14-3-3_pSer127
Modification: (SEP)127
Sequence: TRHIDSSPYS
Construct: TRHID(pSer)SPYS (crude synthetic peptide)
Average holdup BI: 0.14
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 111
P value: 3.53
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 4

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: CRTC2
Fragment: 159-168
Site: 14-3-3_pSer164
Modification: (SEP)164
Sequence: LFRLPSALNR
Construct: LFRLP(pSer)ALNR (crude synthetic peptide)
Average holdup BI: -0.2
Holdup BI standard deviation: 0.26
Immobilized partner concentration (10-6M): 111
P value: 0.82
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: CRTC2
Fragment: 166-175
Site: 14-3-3_pSer171
Modification: (SEP)171
Sequence: LNRTSSDSAL
Construct: LNRTS(pSer)DSAL (crude synthetic peptide)
Average holdup BI: 0.21
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 113
P value: 5.6
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.37
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: CRTC2
Fragment: 166-175
Site: 14-3-3_pSer171
Modification: (SEP)171
Sequence: LNRTSSDSAL
Construct: LNRTS(pSer)DSAL (crude synthetic peptide)
Average holdup BI: 0.21
Holdup BI standard deviation: 0.09
Immobilized partner concentration (10-6M): 142
P value: 2.35
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: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: CRTC2
Fragment: 166-175
Site: 14-3-3_pSer171
Modification: (SEP)171
Sequence: LNRTSSDSAL
Construct: LNRTS(pSer)DSAL (crude synthetic peptide)
Average holdup BI: 0.29
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 111
P value: 7.92
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.57
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: CRTC2
Fragment: 442-451
Site: 14-3-3_pSer447
Modification: (SEP)447
Sequence: ADARRSQQQL
Construct: ADARR(pSer)QQQL (crude synthetic peptide)
Average holdup BI: -0.49
Holdup BI standard deviation: 0.75
Immobilized partner concentration (10-6M): 113
P value: 0.68
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
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: CRTC2
Fragment: 442-451
Site: 14-3-3_pSer447
Modification: (SEP)447
Sequence: ADARRSQQQL
Construct: ADARR(pSer)QQQL (crude synthetic peptide)
Average holdup BI: -0.43
Holdup BI standard deviation: 0.83
Immobilized partner concentration (10-6M): 111
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 10

Protein A

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

Protein B

Protein: CRTC2
Fragment: 487-496
Site: 14-3-3_pSer492
Modification: (SEP)492
Sequence: PYSSPSLVLP
Construct: PYSSP(pSer)LVLP (crude synthetic peptide)
Average holdup BI: 0.12
Holdup BI standard deviation: 0.18
Immobilized partner concentration (10-6M): 113
P value: 0.62
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: CRTC2
Fragment: 487-496
Site: 14-3-3_pSer492
Modification: (SEP)492
Sequence: PYSSPSLVLP
Construct: PYSSP(pSer)LVLP (crude synthetic peptide)
Average holdup BI: 0.03
Holdup BI standard deviation: 0.33
Immobilized partner concentration (10-6M): 142
P value: 0.07
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 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: CRTC2
Fragment: 487-496
Site: 14-3-3_pSer492
Modification: (SEP)492
Sequence: PYSSPSLVLP
Construct: PYSSP(pSer)LVLP (crude synthetic peptide)
Average holdup BI: -0.05
Holdup BI standard deviation: 0.13
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 13

Protein A

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

Protein B

Protein: CRTC2
Fragment: 65-74
Site: 14-3-3_pSer70
Modification: (SEP)70
Sequence: SHYGGSLPNV
Construct: SHYGG(pSer)LPNV (crude synthetic peptide)
Average holdup BI: 0.57
Holdup BI standard deviation: 0.34
Immobilized partner concentration (10-6M): 113
P value: 1.65
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.06
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: CRTC2
Fragment: 65-74
Site: 14-3-3_pSer70
Modification: (SEP)70
Sequence: SHYGGSLPNV
Construct: SHYGG(pSer)LPNV (crude synthetic peptide)
Average holdup BI: 0.53
Holdup BI standard deviation: 0.17
Immobilized partner concentration (10-6M): 142
P value: 3.32
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.9
DOI: 10.1101/2022.12.08.519103
Experimental Details

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