Interaction Details


Details of experiments between O75044 and P62258
Note that the results of all experiments are listed, regardless of the modification states of the fragments.


Experiment series 1

Protein A

Protein: SRGAP2
Fragment: 422-431
Site: 14-3-3_pSer427
Modification: (SEP)427
Sequence: VKSTVSETFM
Construct: VKSTV(pSer)ETFM (crude synthetic peptide)

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.19
Holdup BI standard deviation: 0.08
Immobilized partner concentration (10-6M): 143
P value: 2.69
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.21
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: SRGAP2
Fragment: 849-858
Site: 14-3-3_pSer854
Modification: (SEP)854
Sequence: TFRSDSHGLS
Construct: TFRSD(pSer)HGLS (crude synthetic peptide)

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: -0.08
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 116
P value: 1.51
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 3

Protein A

Protein: SRGAP2
Fragment: 849-858
Site: 14-3-3_pSer854
Modification: (SEP)854
Sequence: TFRSDSHGLS
Construct: TFRSD(pSer)HGLS (crude synthetic peptide)

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: -0.02
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 143
P value: 0.28
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 4

Protein A

Protein: SRGAP2
Fragment: 1009-1018
Site: 14-3-3_pThr1014
Modification: (TPO)1014
Sequence: QRSASTAGDI
Construct: QRSAS(pThr)AGDI (crude synthetic peptide)

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.11
Holdup BI standard deviation: 0.24
Immobilized partner concentration (10-6M): 143
P value: 0.43
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 5

Protein A

Protein: SRGAP2
Fragment: 1065-1074
Site: 14-3-3_pThr1070
Modification: (TPO)1070
Sequence: TDKSCTV---
Construct: PQSTDKSC(pThr)V (crude synthetic peptide)

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Measured dissociation constant (10-6M): <100
Immobilized partner concentration (10-6M): 116
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))