Interaction Details


Details of experiments between P31947 and Q9P2M7
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: CGN
Fragment: 126-135
Site: 14-3-3_pSer131
Modification: (SEP)131
Sequence: LLRSHSQASL
Construct: biotin-ttds-LLRSH(pSer)QASL (HPLC-purified synthetic peptide)
Measured dissociation constant (10-6M): 186
Standard deviation of dissociation constant (10-6M): 13
Average holdup BI: 0
Normalized holdup BI: 0
Normalized holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 0
P value: 0
Experiment method: competitive Fluorescence Polarization
Details of affinity fitting: Competitive binding equation in ProFit
Date of measurement: 2022-02-09
Number of measurements: 3
Measured pKd: 3.73
DOI: 10.1101/2022.12.08.519103

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: CGN
Fragment: 126-135
Site: 14-3-3_pSer131
Modification: (SEP)131
Sequence: LLRSHSQASL
Construct: LLRSH(pSer)QASL (crude synthetic peptide)
Measured dissociation constant (10-6M):
Standard deviation of dissociation constant (10-6M): 0
Average holdup BI: 0.1
Holdup BI standard deviation: 0.04
Normalized holdup BI: 0
Normalized holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 113
P value: 2.88
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: CGN
Fragment: 126-135
Site: 14-3-3_pSer131
Modification: (SEP)131
Sequence: LLRSHSQASL
Construct: LLRSH(pSer)QASL (crude synthetic peptide)
Measured dissociation constant (10-6M):
Standard deviation of dissociation constant (10-6M): 0
Average holdup BI: 0.14
Holdup BI standard deviation: 0.07
Normalized holdup BI: 0
Normalized holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 142
P value: 2.3
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 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: CGN
Fragment: 126-135
Site: 14-3-3_pSer131
Modification: (SEP)131
Sequence: LLRSHSQASL
Construct: LLRSH(pSer)QASL (crude synthetic peptide)
Measured dissociation constant (10-6M):
Standard deviation of dissociation constant (10-6M): 0
Average holdup BI: 0.06
Holdup BI standard deviation: 0.05
Normalized holdup BI: 0
Normalized holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 111
P value: 1.51
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: CGN
Fragment: 144-153
Site: 14-3-3_pSer149
Modification: (SEP)149
Sequence: SNRSNSMLEL
Construct: SNRSN(pSer)MLEL (crude synthetic peptide)
Measured dissociation constant (10-6M):
Standard deviation of dissociation constant (10-6M): 0
Average holdup BI: 0.39
Holdup BI standard deviation: 0.17
Normalized holdup BI: 0
Normalized holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 113
P value: 2.16
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.76
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: CGN
Fragment: 144-153
Site: 14-3-3_pSer149
Modification: (SEP)149
Sequence: SNRSNSMLEL
Construct: SNRSN(pSer)MLEL (crude synthetic peptide)
Measured dissociation constant (10-6M):
Standard deviation of dissociation constant (10-6M): 0
Average holdup BI: 0.25
Holdup BI standard deviation: 0.05
Normalized holdup BI: 0
Normalized holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 142
P value: 4.69
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.38
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: CGN
Fragment: 144-153
Site: 14-3-3_pSer149
Modification: (SEP)149
Sequence: SNRSNSMLEL
Construct: SNRSN(pSer)MLEL (crude synthetic peptide)
Measured dissociation constant (10-6M):
Standard deviation of dissociation constant (10-6M): 0
Average holdup BI: 0.13
Holdup BI standard deviation: 0.41
Normalized holdup BI: 0
Normalized holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 111
P value: 0.27
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: CGN
Fragment: 327-336
Site: 14-3-3_pSer332
Modification: (SEP)332
Sequence: VRRKVSLVLE
Construct: VRRKV(pSer)LVLE (crude synthetic peptide)
Measured dissociation constant (10-6M):
Standard deviation of dissociation constant (10-6M): 0
Average holdup BI: -0.19
Holdup BI standard deviation: 0.29
Normalized holdup BI: 0
Normalized holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 142
P value: 0.71
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: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: CGN
Fragment: 327-336
Site: 14-3-3_pSer332
Modification: (SEP)332
Sequence: VRRKVSLVLE
Construct: VRRKV(pSer)LVLE (crude synthetic peptide)
Measured dissociation constant (10-6M):
Standard deviation of dissociation constant (10-6M): 0
Average holdup BI: 0.07
Holdup BI standard deviation: 0.29
Normalized holdup BI: 0
Normalized holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 111
P value: 0.17
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: CGN
Fragment: 66-75
Site: 14-3-3_pSer71
Modification: (SEP)71
Sequence: EKGGDSFGVQ
Construct: EKGGD(pSer)FGVQ (crude synthetic peptide)
Measured dissociation constant (10-6M):
Standard deviation of dissociation constant (10-6M): 0
Average holdup BI: -0.01
Holdup BI standard deviation: 0.04
Normalized holdup BI: 0
Normalized holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 113
P value: 0.31
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: CGN
Fragment: 66-75
Site: 14-3-3_pSer71
Modification: (SEP)71
Sequence: EKGGDSFGVQ
Construct: EKGGD(pSer)FGVQ (crude synthetic peptide)
Measured dissociation constant (10-6M):
Standard deviation of dissociation constant (10-6M): 0
Average holdup BI: -0.03
Holdup BI standard deviation: 0.03
Normalized holdup BI: 0
Normalized holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 142
P value: 1.26
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: CGN
Fragment: 66-75
Site: 14-3-3_pSer71
Modification: (SEP)71
Sequence: EKGGDSFGVQ
Construct: EKGGD(pSer)FGVQ (crude synthetic peptide)
Measured dissociation constant (10-6M):
Standard deviation of dissociation constant (10-6M): 0
Average holdup BI: 0.02
Holdup BI standard deviation: 0.06
Normalized holdup BI: 0
Normalized holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 111
P value: 0.22
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))