Interaction Details


Details of experiments between Q9P2M7 and P61981
Note that the results of all experiments are listed, regardless of the modification states of the fragments.


Experiment series 1

Protein A

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)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Measured dissociation constant (10-6M): 39
Standard deviation of dissociation constant (10-6M): 3
Experiment method: competitive Fluorescence Polarization
Details of affinity fitting: Competitive binding equation in ProFit
Date of measurement: 2022-01-10
Number of measurements: 3
Measured pKd: 4.41
DOI: 10.1101/2022.12.08.519103

Experiment series 2

Protein A

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)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Measured dissociation constant (10-6M): 47
Standard deviation of dissociation constant (10-6M): 6
Experiment method: competitive Fluorescence Polarization
Details of affinity fitting: Competitive binding equation in ProFit
Date of measurement: 2022-01-18
Number of measurements: 3
Measured pKd: 4.33
DOI: 10.1101/2022.12.08.519103

Experiment series 3

Protein A

Protein: CGN
Fragment: 126-135
Site: 14-3-3_pSer131
Modification: (SEP)131
Sequence: LLRSHSQASL
Construct: LLRSH(pSer)QASL (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.41
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 129
P value: 5.57
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.72
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: CGN
Fragment: 126-135
Site: 14-3-3_pSer131
Modification: (SEP)131
Sequence: LLRSHSQASL
Construct: LLRSH(pSer)QASL (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.41
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 134
P value: 7.61
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.71
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: CGN
Fragment: 144-153
Site: 14-3-3_pSer149
Modification: (SEP)149
Sequence: SNRSNSMLEL
Construct: SNRSN(pSer)MLEL (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.65
Holdup BI standard deviation: 0.11
Immobilized partner concentration (10-6M): 134
P value: 3.69
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.13
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: CGN
Fragment: 144-153
Site: 14-3-3_pSer149
Modification: (SEP)149
Sequence: SNRSNSMLEL
Construct: SNRSN(pSer)MLEL (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.57
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 129
P value: 7.57
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4
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: CGN
Fragment: 327-336
Site: 14-3-3_pSer332
Modification: (SEP)332
Sequence: VRRKVSLVLE
Construct: VRRKV(pSer)LVLE (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.04
Holdup BI standard deviation: 0.2
Immobilized partner concentration (10-6M): 129
P value: 0.15
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 8

Protein A

Protein: CGN
Fragment: 66-75
Site: 14-3-3_pSer71
Modification: (SEP)71
Sequence: EKGGDSFGVQ
Construct: EKGGD(pSer)FGVQ (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: -0.05
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 134
P value: 1.49
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: CGN
Fragment: 66-75
Site: 14-3-3_pSer71
Modification: (SEP)71
Sequence: EKGGDSFGVQ
Construct: EKGGD(pSer)FGVQ (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: -0.07
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 129
P value: 1.78
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 10

Protein A

Protein: CGN
Fragment: 126-135
Site: 14-3-3_pSer131
Modification: (SEP)131
Sequence: LLRSHSQASL
Construct: LLRSH(pSer)QASL (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.3
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 115
P value: 5.83
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.58
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: CGN
Fragment: 144-153
Site: 14-3-3_pSer149
Modification: (SEP)149
Sequence: SNRSNSMLEL
Construct: SNRSN(pSer)MLEL (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.45
Holdup BI standard deviation: 0.32
Immobilized partner concentration (10-6M): 115
P value: 1.48
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.86
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: CGN
Fragment: 327-336
Site: 14-3-3_pSer332
Modification: (SEP)332
Sequence: VRRKVSLVLE
Construct: VRRKV(pSer)LVLE (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.22
Holdup BI standard deviation: 0.24
Immobilized partner concentration (10-6M): 115
P value: 0.84
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: CGN
Fragment: 66-75
Site: 14-3-3_pSer71
Modification: (SEP)71
Sequence: EKGGDSFGVQ
Construct: EKGGD(pSer)FGVQ (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: -0.03
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 115
P value: 0.6
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))