Interaction Details


Details of experiments between Q4KWH8 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: PLCH1
Fragment: 1411-1420
Site: 14-3-3_pSer1416
Modification: (SEP)1416
Sequence: ETLRPSVPEI
Construct: ETLRP(pSer)VPEI (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: 0.2
Immobilized partner concentration (10-6M): 129
P value: 0.13
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 2

Protein A

Protein: PLCH1
Fragment: 1489-1498
Site: 14-3-3_pSer1494
Modification: (SEP)1494
Sequence: PCKSKSLGDL
Construct: PCKSK(pSer)LGDL (crude 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): <10
Immobilized partner concentration (10-6M): 129
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 1
Measured pKd: 5
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: PLCH1
Fragment: 1489-1498
Site: 14-3-3_pSer1494
Modification: (SEP)1494
Sequence: PCKSKSLGDL
Construct: PCKSK(pSer)LGDL (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: -0.49
Holdup BI standard deviation: 2.83
Immobilized partner concentration (10-6M): 134
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 4

Protein A

Protein: PLCH1
Fragment: 1625-1634
Site: 14-3-3_pSer1630
Modification: (SEP)1630
Sequence: AVNRHSTGSY
Construct: AVNRH(pSer)TGSY (crude 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): <100
Immobilized partner concentration (10-6M): 134
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
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 4))

Experiment series 5

Protein A

Protein: PLCH1
Fragment: 918-927
Site: 14-3-3_pSer923
Modification: (SEP)923
Sequence: LRRTASAPAK
Construct: LRRTA(pSer)APAK (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: -0.01
Holdup BI standard deviation: 0.16
Immobilized partner concentration (10-6M): 134
P value: 0.03
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 6

Protein A

Protein: PLCH1
Fragment: 918-927
Site: 14-3-3_pSer923
Modification: (SEP)923
Sequence: LRRTASAPAK
Construct: LRRTA(pSer)APAK (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: 0.13
Immobilized partner concentration (10-6M): 129
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))

Experiment series 7

Protein A

Protein: PLCH1
Fragment: 965-974
Site: 14-3-3_pSer970
Modification: (SEP)970
Sequence: QARPVSMPVD
Construct: QARPV(pSer)MPVD (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: 0.84
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 129
P value: 8.31
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.61
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: PLCH1
Fragment: 965-974
Site: 14-3-3_pSer970
Modification: (SEP)970
Sequence: QARPVSMPVD
Construct: QARPV(pSer)MPVD (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: 0.84
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 134
P value: 6.15
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 9

Protein A

Protein: PLCH1
Fragment: 1315-1324
Site: 14-3-3_pThr1320
Modification: (TPO)1320
Sequence: LPKSPTKGED
Construct: LPKSP(pThr)KGED (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: 0.04
Holdup BI standard deviation: 0.12
Immobilized partner concentration (10-6M): 134
P value: 0.25
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 10

Protein A

Protein: PLCH1
Fragment: 1315-1324
Site: 14-3-3_pThr1320
Modification: (TPO)1320
Sequence: LPKSPTKGED
Construct: LPKSP(pThr)KGED (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: 0.08
Immobilized partner concentration (10-6M): 129
P value: 0.41
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: PLCH1
Fragment: 1515-1524
Site: 14-3-3_pThr1520
Modification: (TPO)1520
Sequence: SKSFVTTGIR
Construct: SKSFV(pThr)TGIR (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: 0.05
Immobilized partner concentration (10-6M): 129
P value: 5.58
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.06
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: PLCH1
Fragment: 1515-1524
Site: 14-3-3_pThr1520
Modification: (TPO)1520
Sequence: SKSFVTTGIR
Construct: SKSFV(pThr)TGIR (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: 0.56
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 134
P value: 5.69
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.98
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 13

Protein A

Protein: PLCH1
Fragment: 1515-1524
Site: 14-3-3_pThr1520
Modification: (TPO)1520
Sequence: SKSFVTTGIR
Construct: biotin-ttds-SKSFV(pThr)TGIR (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): 136
Standard deviation of dissociation constant (10-6M): 24
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: 3.87
DOI: 10.1101/2022.12.08.519103

Experiment series 14

Protein A

Protein: PLCH1
Fragment: 1515-1524
Site: 14-3-3_pThr1520
Modification: (TPO)1520
Sequence: SKSFVTTGIR
Construct: biotin-ttds-SKSFV(pThr)TGIR (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): 159
Standard deviation of dissociation constant (10-6M): 21
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: 3.8
DOI: 10.1101/2022.12.08.519103

Experiment series 15

Protein A

Protein: PLCH1
Fragment: 1411-1420
Site: 14-3-3_pSer1416
Modification: (SEP)1416
Sequence: ETLRPSVPEI
Construct: ETLRP(pSer)VPEI (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: -0.04
Holdup BI standard deviation: 0.12
Immobilized partner concentration (10-6M): 115
P value: 0.28
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 16

Protein A

Protein: PLCH1
Fragment: 918-927
Site: 14-3-3_pSer923
Modification: (SEP)923
Sequence: LRRTASAPAK
Construct: LRRTA(pSer)APAK (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: 0.17
Immobilized partner concentration (10-6M): 115
P value: 0.08
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 17

Protein A

Protein: PLCH1
Fragment: 965-974
Site: 14-3-3_pSer970
Modification: (SEP)970
Sequence: QARPVSMPVD
Construct: QARPV(pSer)MPVD (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: 0.07
Immobilized partner concentration (10-6M): 115
P value: 5.05
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.44
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: PLCH1
Fragment: 1315-1324
Site: 14-3-3_pThr1320
Modification: (TPO)1320
Sequence: LPKSPTKGED
Construct: LPKSP(pThr)KGED (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: -0.05
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 115
P value: 0.82
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: PLCH1
Fragment: 1515-1524
Site: 14-3-3_pThr1520
Modification: (TPO)1520
Sequence: SKSFVTTGIR
Construct: SKSFV(pThr)TGIR (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: 0.71
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 115
P value: 7.79
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.34
DOI: 10.1101/2022.12.08.519103
Experimental Details

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