Interaction Details


Details of experiments between P31947 and Q86X27
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: RALGPS2
Fragment: 291-300
Site: 14-3-3_pSer296
Modification: (SEP)296
Sequence: PRSAASREDL
Construct: PRSAA(pSer)REDL (crude synthetic peptide)
Average holdup BI: 0.01
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 113
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: RALGPS2
Fragment: 291-300
Site: 14-3-3_pSer296
Modification: (SEP)296
Sequence: PRSAASREDL
Construct: PRSAA(pSer)REDL (crude synthetic peptide)
Average holdup BI: -0.01
Holdup BI standard deviation: 0.06
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 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: RALGPS2
Fragment: 291-300
Site: 14-3-3_pSer296
Modification: (SEP)296
Sequence: PRSAASREDL
Construct: PRSAA(pSer)REDL (crude synthetic peptide)
Average holdup BI: -0.1
Holdup BI standard deviation: 0.15
Immobilized partner concentration (10-6M): 111
P value: 0.72
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: RALGPS2
Fragment: 417-426
Site: 14-3-3_pSer422
Modification: (SEP)422
Sequence: NRLYHSLGPV
Construct: NRLYH(pSer)LGPV (crude synthetic peptide)
Average holdup BI: 0.68
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 142
P value: 5.58
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.16
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
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: RALGPS2
Fragment: 417-426
Site: 14-3-3_pSer422
Modification: (SEP)422
Sequence: NRLYHSLGPV
Construct: NRLYH(pSer)LGPV (crude synthetic peptide)
Average holdup BI: 0.56
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 113
P value: 4.69
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.04
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
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: RALGPS2
Fragment: 417-426
Site: 14-3-3_pSer422
Modification: (SEP)422
Sequence: NRLYHSLGPV
Construct: NRLYH(pSer)LGPV (crude synthetic peptide)
Average holdup BI: 0.65
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 111
P value: 5.66
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.22
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
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: RALGPS2
Fragment: 356-365
Site: 14-3-3_pThr361
Modification: (TPO)361
Sequence: EFKSATFPNA
Construct: EFKSA(pThr)FPNA (crude synthetic peptide)
Average holdup BI: 0.9
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 142
P value: 8.78
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.78
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

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: RALGPS2
Fragment: 356-365
Site: 14-3-3_pThr361
Modification: (TPO)361
Sequence: EFKSATFPNA
Construct: EFKSA(pThr)FPNA (crude synthetic peptide)
Average holdup BI: 0.87
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 113
P value: 10.11
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.76
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: RALGPS2
Fragment: 356-365
Site: 14-3-3_pThr361
Modification: (TPO)361
Sequence: EFKSATFPNA
Construct: EFKSA(pThr)FPNA (crude synthetic peptide)
Average holdup BI: 0.84
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 111
P value: 8.81
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.68
DOI: 10.1101/2022.12.08.519103
Experimental Details

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