Interaction Details


Details of experiments between Q9P2M7 and P63104
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: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta
Measured dissociation constant (10-6M): 96
Standard deviation of dissociation constant (10-6M): 30
Experiment method: competitive Fluorescence Polarization
Details of affinity fitting: Competitive binding equation in ProFit
Date of measurement: 2022-02-10
Number of measurements: 3
Measured pKd: 4.02
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: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta
Measured dissociation constant (10-6M): 105
Standard deviation of dissociation constant (10-6M): 2
Experiment method: competitive Fluorescence Polarization
Details of affinity fitting: Competitive binding equation in ProFit
Date of measurement: 2022-01-19
Number of measurements: 3
Measured pKd: 3.98
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: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta
Average holdup BI: 0.24
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 132
P value: 3.74
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.37
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: 144-153
Site: 14-3-3_pSer149
Modification: (SEP)149
Sequence: SNRSNSMLEL
Construct: SNRSN(pSer)MLEL (crude synthetic peptide)

Protein B

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta
Average holdup BI: 0.38
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 132
P value: 5.09
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.67
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: CGN
Fragment: 327-336
Site: 14-3-3_pSer332
Modification: (SEP)332
Sequence: VRRKVSLVLE
Construct: VRRKV(pSer)LVLE (crude synthetic peptide)

Protein B

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta
Average holdup BI: 0.01
Holdup BI standard deviation: 0.21
Immobilized partner concentration (10-6M): 132
P value: 0.04
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 6

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: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta
Average holdup BI: -0.04
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 132
P value: 1.3
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))