Interaction Details


Details of experiments between P61981 and Q86X27
Note that the results of all experiments are listed, regardless of the modification states of the fragments.


Experiment series 1

Protein A

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

Protein B

Protein: RALGPS2
Fragment: 291-300
Site: 14-3-3_pSer296
Modification: (SEP)296
Sequence: PRSAASREDL
Construct: PRSAA(pSer)REDL (crude synthetic peptide)
Average holdup BI: 0.03
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 134
P value: 0.57
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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

Protein B

Protein: RALGPS2
Fragment: 291-300
Site: 14-3-3_pSer296
Modification: (SEP)296
Sequence: PRSAASREDL
Construct: PRSAA(pSer)REDL (crude synthetic peptide)
Average holdup BI: -0.03
Holdup BI standard deviation: 0.08
Immobilized partner concentration (10-6M): 129
P value: 0.39
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: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

Protein B

Protein: RALGPS2
Fragment: 291-300
Site: 14-3-3_pSer296
Modification: (SEP)296
Sequence: PRSAASREDL
Construct: PRSAA(pSer)REDL (crude synthetic peptide)
Average holdup BI: -0.06
Holdup BI standard deviation: 0.15
Immobilized partner concentration (10-6M): 115
P value: 0.34
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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

Protein B

Protein: RALGPS2
Fragment: 417-426
Site: 14-3-3_pSer422
Modification: (SEP)422
Sequence: NRLYHSLGPV
Construct: NRLYH(pSer)LGPV (crude synthetic peptide)
Average holdup BI: 0.95
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 129
P value: 7.04
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 5.17
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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

Protein B

Protein: RALGPS2
Fragment: 417-426
Site: 14-3-3_pSer422
Modification: (SEP)422
Sequence: NRLYHSLGPV
Construct: NRLYH(pSer)LGPV (crude synthetic peptide)
Average holdup BI: 0.35
Holdup BI standard deviation: 1.07
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 6

Protein A

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

Protein B

Protein: RALGPS2
Fragment: 417-426
Site: 14-3-3_pSer422
Modification: (SEP)422
Sequence: NRLYHSLGPV
Construct: NRLYH(pSer)LGPV (crude synthetic peptide)
Measured dissociation constant (10-6M): <10
Immobilized partner concentration (10-6M): 115
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
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 4))

Experiment series 7

Protein A

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

Protein B

Protein: RALGPS2
Fragment: 356-365
Site: 14-3-3_pThr361
Modification: (TPO)361
Sequence: EFKSATFPNA
Construct: EFKSA(pThr)FPNA (crude synthetic peptide)
Average holdup BI: 0.94
Holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 129
P value: 9.2
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 5.1
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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

Protein B

Protein: RALGPS2
Fragment: 356-365
Site: 14-3-3_pThr361
Modification: (TPO)361
Sequence: EFKSATFPNA
Construct: EFKSA(pThr)FPNA (crude synthetic peptide)
Average holdup BI: 0.94
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 134
P value: 10.2
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 5.05
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: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma

Protein B

Protein: RALGPS2
Fragment: 356-365
Site: 14-3-3_pThr361
Modification: (TPO)361
Sequence: EFKSATFPNA
Construct: EFKSA(pThr)FPNA (crude synthetic peptide)
Average holdup BI: 0.91
Holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 115
P value: 9.23
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.98
DOI: 10.1101/2022.12.08.519103
Experimental Details

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