Interaction Details


Details of experiments between Q5VZ89 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: DENND4C
Fragment: 1094-1103
Site: 14-3-3_pSer1099
Modification: (SEP)1099
Sequence: IARTHSFENV
Construct: IARTH(pSer)FENV (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: 0.09
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 132
P value: 4.07
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 2

Protein A

Protein: DENND4C
Fragment: 1179-1188
Site: 14-3-3_pSer1184
Modification: (SEP)1184
Sequence: RNKRSSLYGI
Construct: RNKRS(pSer)LYGI (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: 0.28
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 132
P value: 3.89
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.48
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: DENND4C
Fragment: 1273-1282
Site: 14-3-3_pSer1278
Modification: (SEP)1278
Sequence: LERRSSLPLD
Construct: LERRS(pSer)LPLD (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: 0.04
Immobilized partner concentration (10-6M): 132
P value: 5.94
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 4

Protein A

Protein: DENND4C
Fragment: 1320-1329
Site: 14-3-3_pSer1325
Modification: (SEP)1325
Sequence: KERSTSLSAL
Construct: KERST(pSer)LSAL (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: 0.77
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 132
P value: 7.25
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.39
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: DENND4C
Fragment: 1320-1329
Site: 14-3-3_pSer1325
Modification: (SEP)1325
Sequence: KERSTSLSAL
Construct: biotin-ttds-KERST(pSer)LSAL (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): 58
Standard deviation of dissociation constant (10-6M): 0
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: 4.24
DOI: 10.1101/2022.12.08.519103

Experiment series 6

Protein A

Protein: DENND4C
Fragment: 1320-1329
Site: 14-3-3_pSer1325
Modification: (SEP)1325
Sequence: KERSTSLSAL
Construct: biotin-ttds-KERST(pSer)LSAL (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): 64
Standard deviation of dissociation constant (10-6M): 13
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: 4.19
DOI: 10.1101/2022.12.08.519103

Experiment series 7

Protein A

Protein: DENND4C
Fragment: 1341-1350
Site: 14-3-3_pSer1346
Modification: (SEP)1346
Sequence: VVNSLSGLKL
Construct: VVNSL(pSer)GLKL (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: -0.11
Holdup BI standard deviation: 0.23
Immobilized partner concentration (10-6M): 132
P value: 0.55
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: DENND4C
Fragment: 1618-1627
Site: 14-3-3_pSer1623
Modification: (SEP)1623
Sequence: TMKISSVPNS
Construct: TMKIS(pSer)VPNS (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: 0.42
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 132
P value: 7.12
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.74
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: DENND4C
Fragment: 1635-1644
Site: 14-3-3_pSer1640
Modification: (SEP)1640
Sequence: LTRSHSVGGP
Construct: LTRSH(pSer)VGGP (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: 0.64
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 132
P value: 8.04
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.12
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: DENND4C
Fragment: 1657-1666
Site: 14-3-3_pSer1662
Modification: (SEP)1662
Sequence: GISTVSLPNS
Construct: GISTV(pSer)LPNS (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: 0.72
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 132
P value: 8.42
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 11

Protein A

Protein: DENND4C
Fragment: 1661-1670
Site: 14-3-3_pSer1666
Modification: (SEP)1666
Sequence: VSLPNSLQEV
Construct: VSLPN(pSer)LQEV (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: 0.02
Holdup BI standard deviation: 0.17
Immobilized partner concentration (10-6M): 132
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 12

Protein A

Protein: DENND4C
Fragment: 948-957
Site: 14-3-3_pSer953
Modification: (SEP)953
Sequence: LIRLESIDNH
Construct: LIRLE(pSer)IDNH (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: -0.08
Holdup BI standard deviation: 0.09
Immobilized partner concentration (10-6M): 132
P value: 1.13
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 13

Protein A

Protein: DENND4C
Fragment: 960-969
Site: 14-3-3_pSer965
Modification: (SEP)965
Sequence: IVAKHSQPSP
Construct: IVAKH(pSer)QPSP (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: 0.11
Immobilized partner concentration (10-6M): 132
P value: 0.3
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 14

Protein A

Protein: DENND4C
Fragment: 984-993
Site: 14-3-3_pSer989
Modification: (SEP)989
Sequence: IVKVPSGIFD
Construct: IVKVP(pSer)GIFD (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: -0.36
Holdup BI standard deviation: 0.35
Immobilized partner concentration (10-6M): 132
P value: 1.77
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))