Interaction Details


Details of experiments between Q5T5U3 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: ARHGAP21
Fragment: 5-14
Site: 14-3-3_pSer10
Modification: (SEP)10
Sequence: RRTGLSEGDG
Construct: RRTGL(pSer)EGDG (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 2

Protein A

Protein: ARHGAP21
Fragment: 1487-1496
Site: 14-3-3_pSer1492
Modification: (SEP)1492
Sequence: KDEKISLGKE
Construct: KDEKI(pSer)LGKE (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
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 134
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 3

Protein A

Protein: ARHGAP21
Fragment: 1487-1496
Site: 14-3-3_pSer1492
Modification: (SEP)1492
Sequence: KDEKISLGKE
Construct: KDEKI(pSer)LGKE (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: 0.07
Immobilized partner concentration (10-6M): 129
P value: 0.51
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 4

Protein A

Protein: ARHGAP21
Fragment: 1492-1501
Site: 14-3-3_pSer1497
Modification: (SEP)1497
Sequence: SLGKESTPSE
Construct: SLGKE(pSer)TPSE (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: 0.05
Immobilized partner concentration (10-6M): 134
P value: 2.21
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 5

Protein A

Protein: ARHGAP21
Fragment: 1492-1501
Site: 14-3-3_pSer1497
Modification: (SEP)1497
Sequence: SLGKESTPSE
Construct: SLGKE(pSer)TPSE (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: -0.08
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 129
P value: 1.81
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 6

Protein A

Protein: ARHGAP21
Fragment: 1712-1721
Site: 14-3-3_pSer1717
Modification: (SEP)1717
Sequence: KSDSGSLGDA
Construct: KSDSG(pSer)LGDA (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: -0.1
Holdup BI standard deviation: 0.3
Immobilized partner concentration (10-6M): 134
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 7

Protein A

Protein: ARHGAP21
Fragment: 1737-1746
Site: 14-3-3_pSer1742
Modification: (SEP)1742
Sequence: MKKGKSTGSL
Construct: MKKGK(pSer)TGSL (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
Holdup BI standard deviation: 0.24
Immobilized partner concentration (10-6M): 134
P value: 0.01
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 8

Protein A

Protein: ARHGAP21
Fragment: 1737-1746
Site: 14-3-3_pSer1742
Modification: (SEP)1742
Sequence: MKKGKSTGSL
Construct: MKKGK(pSer)TGSL (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: 0.1
Immobilized partner concentration (10-6M): 129
P value: 0.4
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: ARHGAP21
Fragment: 228-237
Site: 14-3-3_pSer233
Modification: (SEP)233
Sequence: SKQQTSTPVL
Construct: SKQQT(pSer)TPVL (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: 0.42
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 129
P value: 10.17
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.75
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: ARHGAP21
Fragment: 228-237
Site: 14-3-3_pSer233
Modification: (SEP)233
Sequence: SKQQTSTPVL
Construct: SKQQT(pSer)TPVL (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: 0.41
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 134
P value: 6.8
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.71
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 11

Protein A

Protein: ARHGAP21
Fragment: 454-463
Site: 14-3-3_pSer459
Modification: (SEP)459
Sequence: RQRSVSQERL
Construct: RQRSV(pSer)QERL (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: 0.42
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 129
P value: 5.06
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.75
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: ARHGAP21
Fragment: 454-463
Site: 14-3-3_pSer459
Modification: (SEP)459
Sequence: RQRSVSQERL
Construct: RQRSV(pSer)QERL (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: 0.25
Holdup BI standard deviation: 0.13
Immobilized partner concentration (10-6M): 134
P value: 1.93
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 13

Protein A

Protein: ARHGAP21
Fragment: 52-61
Site: 14-3-3_pSer57
Modification: (SEP)57
Sequence: TLKRTSQGFG
Construct: TLKRT(pSer)QGFG (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: 0.03
Immobilized partner concentration (10-6M): 134
P value: 0.99
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 14

Protein A

Protein: ARHGAP21
Fragment: 52-61
Site: 14-3-3_pSer57
Modification: (SEP)57
Sequence: TLKRTSQGFG
Construct: TLKRT(pSer)QGFG (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: 0.05
Immobilized partner concentration (10-6M): 129
P value: 0.83
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 15

Protein A

Protein: ARHGAP21
Fragment: 607-616
Site: 14-3-3_pSer612
Modification: (SEP)612
Sequence: LPRGISQDRS
Construct: LPRGI(pSer)QDRS (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: 0.09
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 134
P value: 2.28
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: ARHGAP21
Fragment: 607-616
Site: 14-3-3_pSer612
Modification: (SEP)612
Sequence: LPRGISQDRS
Construct: LPRGI(pSer)QDRS (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: 0.05
Immobilized partner concentration (10-6M): 129
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))

Experiment series 17

Protein A

Protein: ARHGAP21
Fragment: 666-675
Site: 14-3-3_pSer671
Modification: (SEP)671
Sequence: WVRTDSAPDQ
Construct: WVRTD(pSer)APDQ (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: 0.21
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 134
P value: 4.13
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 18

Protein A

Protein: ARHGAP21
Fragment: 666-675
Site: 14-3-3_pSer671
Modification: (SEP)671
Sequence: WVRTDSAPDQ
Construct: WVRTD(pSer)APDQ (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: 0.04
Immobilized partner concentration (10-6M): 129
P value: 2.78
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 19

Protein A

Protein: ARHGAP21
Fragment: 5-14
Site: 14-3-3_pSer10
Modification: (SEP)10
Sequence: RRTGLSEGDG
Construct: RRTGL(pSer)EGDG (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: -0.07
Holdup BI standard deviation: 0.64
Immobilized partner concentration (10-6M): 115
P value: 0.06
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 20

Protein A

Protein: ARHGAP21
Fragment: 1487-1496
Site: 14-3-3_pSer1492
Modification: (SEP)1492
Sequence: KDEKISLGKE
Construct: KDEKI(pSer)LGKE (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: -0.06
Holdup BI standard deviation: 0.16
Immobilized partner concentration (10-6M): 115
P value: 0.32
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: ARHGAP21
Fragment: 1492-1501
Site: 14-3-3_pSer1497
Modification: (SEP)1497
Sequence: SLGKESTPSE
Construct: SLGKE(pSer)TPSE (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: -0.09
Holdup BI standard deviation: 0.08
Immobilized partner concentration (10-6M): 115
P value: 1.33
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 22

Protein A

Protein: ARHGAP21
Fragment: 1708-1717
Site: 14-3-3_pSer1713
Modification: (SEP)1713
Sequence: SARFKSDSGS
Construct: SARFK(pSer)DSGS (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: -0.19
Holdup BI standard deviation: 1.6
Immobilized partner concentration (10-6M): 115
P value: 0.09
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: ARHGAP21
Fragment: 228-237
Site: 14-3-3_pSer233
Modification: (SEP)233
Sequence: SKQQTSTPVL
Construct: SKQQT(pSer)TPVL (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: 0.32
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 115
P value: 4.97
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.62
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 24

Protein A

Protein: ARHGAP21
Fragment: 454-463
Site: 14-3-3_pSer459
Modification: (SEP)459
Sequence: RQRSVSQERL
Construct: RQRSV(pSer)QERL (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: 0.11
Holdup BI standard deviation: 0.2
Immobilized partner concentration (10-6M): 115
P value: 0.52
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 25

Protein A

Protein: ARHGAP21
Fragment: 52-61
Site: 14-3-3_pSer57
Modification: (SEP)57
Sequence: TLKRTSQGFG
Construct: TLKRT(pSer)QGFG (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: -0.01
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 115
P value: 0.07
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: ARHGAP21
Fragment: 607-616
Site: 14-3-3_pSer612
Modification: (SEP)612
Sequence: LPRGISQDRS
Construct: LPRGI(pSer)QDRS (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: -0.06
Holdup BI standard deviation: 0.12
Immobilized partner concentration (10-6M): 115
P value: 0.51
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: ARHGAP21
Fragment: 666-675
Site: 14-3-3_pSer671
Modification: (SEP)671
Sequence: WVRTDSAPDQ
Construct: WVRTD(pSer)APDQ (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: 0.14
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 115
P value: 2.11
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))