Interaction Details


Details of experiments between P63104 and Q9UQC2
Note that the results of all experiments are listed, regardless of the modification states of the fragments.


Experiment series 1

Protein A

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta

Protein B

Protein: GAB2
Fragment: 128-137
Site: 14-3-3_pSer133
Modification: (SEP)133
Sequence: LRNVSSAGHG
Construct: LRNVS(pSer)AGHG (crude synthetic peptide)
Measured dissociation constant (10-6M): <100
Immobilized partner concentration (10-6M): 132
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 1
Measured pKd: 4
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 2

Protein A

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta

Protein B

Protein: GAB2
Fragment: 205-214
Site: 14-3-3_pSer210
Modification: (SEP)210
Sequence: NARSASFSQG
Construct: NARSA(pSer)FSQG (crude synthetic peptide)
Average holdup BI: 0.37
Holdup BI standard deviation: 0.14
Immobilized partner concentration (10-6M): 132
P value: 2.96
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.65
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: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta

Protein B

Protein: GAB2
Fragment: 207-216
Site: 14-3-3_pSer212
Modification: (SEP)212
Sequence: RSASFSQGTR
Construct: biotin-ttds-RSASF(pSer)QGTR (HPLC-purified synthetic peptide)
Measured dissociation constant (10-6M): 166
Standard deviation of dissociation constant (10-6M): 53
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: 3.78
DOI: 10.1101/2022.12.08.519103

Experiment series 4

Protein A

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta

Protein B

Protein: GAB2
Fragment: 207-216
Site: 14-3-3_pSer212
Modification: (SEP)212
Sequence: RSASFSQGTR
Construct: biotin-ttds-RSASF(pSer)QGTR (HPLC-purified synthetic peptide)
Measured dissociation constant (10-6M): 195
Standard deviation of dissociation constant (10-6M): 7
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: 3.71
DOI: 10.1101/2022.12.08.519103

Experiment series 5

Protein A

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta

Protein B

Protein: GAB2
Fragment: 207-216
Site: 14-3-3_pSer212
Modification: (SEP)212
Sequence: RSASFSQGTR
Construct: RSASF(pSer)QGTR (crude synthetic peptide)
Average holdup BI: 0.39
Holdup BI standard deviation: 0.08
Immobilized partner concentration (10-6M): 132
P value: 4.34
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.68
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: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta

Protein B

Protein: GAB2
Fragment: 259-268
Site: 14-3-3_pSer264
Modification: (SEP)264
Sequence: TEFRDSTYDL
Construct: TEFRD(pSer)TYDL (crude synthetic peptide)
Average holdup BI: -0.07
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 132
P value: 1.18
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: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta

Protein B

Protein: GAB2
Fragment: 317-326
Site: 14-3-3_pSer322
Modification: (SEP)322
Sequence: PATPLSAYQI
Construct: PATPL(pSer)AYQI (crude synthetic peptide)
Average holdup BI: 0.05
Holdup BI standard deviation: 0.13
Immobilized partner concentration (10-6M): 132
P value: 0.31
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: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta

Protein B

Protein: GAB2
Fragment: 467-476
Site: 14-3-3_pSer472
Modification: (SEP)472
Sequence: RAGDNSQSVY
Construct: RAGDN(pSer)QSVY (crude synthetic peptide)
Average holdup BI: -0.03
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 132
P value: 0.43
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: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta

Protein B

Protein: GAB2
Fragment: 380-389
Site: 14-3-3_pThr385
Modification: (TPO)385
Sequence: RSVAATIPRR
Construct: RSVAA(pThr)IPRR (crude synthetic peptide)
Average holdup BI: 0.72
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 132
P value: 7.89
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.28
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: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta

Protein B

Protein: GAB2
Fragment: 386-395
Site: 14-3-3_pThr391
Modification: (TPO)391
Sequence: IPRRNTLPAM
Construct: IPRRN(pThr)LPAM (crude synthetic peptide)
Average holdup BI: 0.96
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 132
P value: 6.9
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 5.22
DOI: 10.1101/2022.12.08.519103
Experimental Details

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