Interaction Details


Details of experiments between Q5VZ89 and Q04917
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: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta
Average holdup BI: 0.12
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 130
P value: 2.76
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: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta
Average holdup BI: 0.43
Holdup BI standard deviation: 0.1
Immobilized partner concentration (10-6M): 130
P value: 4.13
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.76
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: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta
Average holdup BI: 0.54
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 130
P value: 6.73
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.95
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: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta
Average holdup BI: 0.82
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 130
P value: 6.82
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.54
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: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta
Measured dissociation constant (10-6M): 31
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: 4.51
DOI: 10.1101/2022.12.08.519103

Experiment series 6

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: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta
Average holdup BI: 0.15
Holdup BI standard deviation: 0.25
Immobilized partner concentration (10-6M): 130
P value: 0.53
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: DENND4C
Fragment: 1618-1627
Site: 14-3-3_pSer1623
Modification: (SEP)1623
Sequence: TMKISSVPNS
Construct: TMKIS(pSer)VPNS (crude synthetic peptide)

Protein B

Protein: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta
Average holdup BI: 0.47
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 130
P value: 6.52
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.83
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: 1635-1644
Site: 14-3-3_pSer1640
Modification: (SEP)1640
Sequence: LTRSHSVGGP
Construct: LTRSH(pSer)VGGP (crude synthetic peptide)

Protein B

Protein: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta
Average holdup BI: 0.7
Holdup BI standard deviation: 0.12
Immobilized partner concentration (10-6M): 130
P value: 4.97
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.25
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: 1657-1666
Site: 14-3-3_pSer1662
Modification: (SEP)1662
Sequence: GISTVSLPNS
Construct: GISTV(pSer)LPNS (crude synthetic peptide)

Protein B

Protein: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta
Average holdup BI: 0.76
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 130
P value: 7
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.38
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: 1661-1670
Site: 14-3-3_pSer1666
Modification: (SEP)1666
Sequence: VSLPNSLQEV
Construct: VSLPN(pSer)LQEV (crude synthetic peptide)

Protein B

Protein: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta
Average holdup BI: 0.19
Holdup BI standard deviation: 0.18
Immobilized partner concentration (10-6M): 130
P value: 1.07
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 11

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: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta
Average holdup BI: -0.02
Holdup BI standard deviation: 0.11
Immobilized partner concentration (10-6M): 130
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: 960-969
Site: 14-3-3_pSer965
Modification: (SEP)965
Sequence: IVAKHSQPSP
Construct: IVAKH(pSer)QPSP (crude synthetic peptide)

Protein B

Protein: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta
Average holdup BI: 0.03
Holdup BI standard deviation: 0.1
Immobilized partner concentration (10-6M): 130
P value: 0.23
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: 984-993
Site: 14-3-3_pSer989
Modification: (SEP)989
Sequence: IVKVPSGIFD
Construct: IVKVP(pSer)GIFD (crude synthetic peptide)

Protein B

Protein: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta
Average holdup BI: 0.06
Holdup BI standard deviation: 0.39
Immobilized partner concentration (10-6M): 130
P value: 0.12
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))