Interaction Details


Details of experiments between Q5VZ89 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: DENND4C
Fragment: 1094-1103
Site: 14-3-3_pSer1099
Modification: (SEP)1099
Sequence: IARTHSFENV
Construct: IARTH(pSer)FENV (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: 0.19
Holdup BI standard deviation: 0.08
Immobilized partner concentration (10-6M): 134
P value: 2.27
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 2

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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.14
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 129
P value: 3.78
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 3

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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.44
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 134
P value: 4.3
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.76
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 4

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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.43
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 129
P value: 5.27
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 5

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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.59
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 129
P value: 6.95
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.04
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: DENND4C
Fragment: 1273-1282
Site: 14-3-3_pSer1278
Modification: (SEP)1278
Sequence: LERRSSLPLD
Construct: LERRS(pSer)LPLD (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: 0.53
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 134
P value: 7.19
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.92
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 7

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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.87
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 129
P value: 7.84
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.7
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: 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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Measured dissociation constant (10-6M): 23
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: 2022-01-10
Number of measurements: 3
Measured pKd: 4.65
DOI: 10.1101/2022.12.08.519103

Experiment series 9

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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.85
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 134
P value: 7.74
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.63
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 10

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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Measured dissociation constant (10-6M): 24
Standard deviation of dissociation constant (10-6M): 2.5
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: 4.61
DOI: 10.1101/2022.12.08.519103

Experiment series 11

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: 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: 0.3
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 12

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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.66
Holdup BI standard deviation: 0.08
Immobilized partner concentration (10-6M): 129
P value: 6.05
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.18
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: 1618-1627
Site: 14-3-3_pSer1623
Modification: (SEP)1623
Sequence: TMKISSVPNS
Construct: TMKIS(pSer)VPNS (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: 0.66
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 134
P value: 5.1
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.17
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 14

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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.77
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 129
P value: 7.69
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.42
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 15

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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.76
Holdup BI standard deviation: 0.09
Immobilized partner concentration (10-6M): 134
P value: 4.76
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.38
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 16

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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.87
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 129
P value: 9.31
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.71
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 17

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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.83
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 134
P value: 8.44
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.55
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 18

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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.13
Holdup BI standard deviation: 0.43
Immobilized partner concentration (10-6M): 134
P value: 0.22
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 19

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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.2
Holdup BI standard deviation: 0.16
Immobilized partner concentration (10-6M): 129
P value: 1.28
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 20

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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: -0.29
Holdup BI standard deviation: 0.1
Immobilized partner concentration (10-6M): 134
P value: 3.14
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 21

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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: -0.11
Holdup BI standard deviation: 0.13
Immobilized partner concentration (10-6M): 129
P value: 0.98
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 22

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: 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: 0.15
Immobilized partner concentration (10-6M): 134
P value: 0.43
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 23

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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.03
Holdup BI standard deviation: 0.11
Immobilized partner concentration (10-6M): 129
P value: 0.19
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 24

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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: -0.15
Holdup BI standard deviation: 0.42
Immobilized partner concentration (10-6M): 129
P value: 0.35
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 25

Protein A

Protein: DENND4C
Fragment: 1041-1050
Site: 14-3-3_pSer1046
Modification: (SEP)1046
Sequence: PERSCSFSSE
Construct: PERSC(pSer)FSSE (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: -0.15
Holdup BI standard deviation: 0.3
Immobilized partner concentration (10-6M): 115
P value: 0.5
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 26

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: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.2
Holdup BI standard deviation: 0.11
Immobilized partner concentration (10-6M): 115
P value: 1.71
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 27

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: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.46
Holdup BI standard deviation: 0.13
Immobilized partner concentration (10-6M): 115
P value: 2.98
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.88
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 28

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: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.47
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 115
P value: 5.21
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.9
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 29

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: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.81
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 115
P value: 8.88
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.58
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 30

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: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.24
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 115
P value: 3.3
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 31

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: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.75
Holdup BI standard deviation: 0.09
Immobilized partner concentration (10-6M): 115
P value: 5.27
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.43
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 32

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: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.81
Holdup BI standard deviation: 0.08
Immobilized partner concentration (10-6M): 115
P value: 1.9
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.59
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 33

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: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.78
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 115
P value: 7.14
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.5
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 34

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: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.53
Holdup BI standard deviation: 0.29
Immobilized partner concentration (10-6M): 115
P value: 1.6
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.01
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 35

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: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 115
P value: 0
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 36

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: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: -0.02
Holdup BI standard deviation: 0.1
Immobilized partner concentration (10-6M): 115
P value: 0.14
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 37

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: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.02
Holdup BI standard deviation: 0.1
Immobilized partner concentration (10-6M): 115
P value: 0.17
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))