Interaction Details


Details of experiments between Q9UJ41 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: RABGEF1
Fragment: 101-110
Site: 14-3-3_pSer106
Modification: (SEP)106
Sequence: VKKFFSASSR
Construct: VKKFF(pSer)ASSR (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: 0.47
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 116
P value: 5.78
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 2

Protein A

Protein: RABGEF1
Fragment: 101-110
Site: 14-3-3_pSer106
Modification: (SEP)106
Sequence: VKKFFSASSR
Construct: VKKFF(pSer)ASSR (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: 0.48
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 143
P value: 5.52
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.81
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: RABGEF1
Fragment: 108-117
Site: 14-3-3_pSer113
Modification: (SEP)113
Sequence: SSRVGSKKEI
Construct: SSRVG(pSer)KKEI (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: -0.04
Holdup BI standard deviation: 0.13
Immobilized partner concentration (10-6M): 143
P value: 0.23
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: RABGEF1
Fragment: 127-136
Site: 14-3-3_pSer132
Modification: (SEP)132
Sequence: INRQTSIETD
Construct: INRQT(pSer)IETD (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: -0.05
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 116
P value: 1.31
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: RABGEF1
Fragment: 127-136
Site: 14-3-3_pSer132
Modification: (SEP)132
Sequence: INRQTSIETD
Construct: INRQT(pSer)IETD (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: -0.04
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 143
P value: 0.73
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: RABGEF1
Fragment: 372-381
Site: 14-3-3_pSer377
Modification: (SEP)377
Sequence: QSLNLSQEDF
Construct: QSLNL(pSer)QEDF (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: 0.08
Immobilized partner concentration (10-6M): 116
P value: 0.98
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: RABGEF1
Fragment: 372-381
Site: 14-3-3_pSer377
Modification: (SEP)377
Sequence: QSLNLSQEDF
Construct: QSLNL(pSer)QEDF (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: -0.17
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 143
P value: 3.43
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 8

Protein A

Protein: RABGEF1
Fragment: 384-393
Site: 14-3-3_pThr389
Modification: (TPO)389
Sequence: YMSGQTSPRK
Construct: YMSGQ(pThr)SPRK (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: 0.24
Holdup BI standard deviation: 0.22
Immobilized partner concentration (10-6M): 116
P value: 0.69
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 9

Protein A

Protein: RABGEF1
Fragment: 384-393
Site: 14-3-3_pThr389
Modification: (TPO)389
Sequence: YMSGQTSPRK
Construct: YMSGQ(pThr)SPRK (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: -0.01
Holdup BI standard deviation: 0.63
Immobilized partner concentration (10-6M): 143
P value: 0.01
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))