Interaction Details


Details of experiments between P31947 and Q9UJ41
Note that the results of all experiments are listed, regardless of the modification states of the fragments.


Experiment series 1

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: RABGEF1
Fragment: 101-110
Site: 14-3-3_pSer106
Modification: (SEP)106
Sequence: VKKFFSASSR
Construct: VKKFF(pSer)ASSR (crude synthetic peptide)
Average holdup BI: 0.54
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 113
P value: 7.07
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.02
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: RABGEF1
Fragment: 101-110
Site: 14-3-3_pSer106
Modification: (SEP)106
Sequence: VKKFFSASSR
Construct: VKKFF(pSer)ASSR (crude synthetic peptide)
Average holdup BI: 0.58
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 142
P value: 6.19
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.98
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: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: RABGEF1
Fragment: 101-110
Site: 14-3-3_pSer106
Modification: (SEP)106
Sequence: VKKFFSASSR
Construct: VKKFF(pSer)ASSR (crude synthetic peptide)
Average holdup BI: 0.56
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 111
P value: 4.96
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.05
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: RABGEF1
Fragment: 108-117
Site: 14-3-3_pSer113
Modification: (SEP)113
Sequence: SSRVGSKKEI
Construct: SSRVG(pSer)KKEI (crude synthetic peptide)
Average holdup BI: -0.08
Holdup BI standard deviation: 0.11
Immobilized partner concentration (10-6M): 142
P value: 0.75
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: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: RABGEF1
Fragment: 108-117
Site: 14-3-3_pSer113
Modification: (SEP)113
Sequence: SSRVGSKKEI
Construct: SSRVG(pSer)KKEI (crude synthetic peptide)
Average holdup BI: 0.11
Holdup BI standard deviation: 0.3
Immobilized partner concentration (10-6M): 111
P value: 0.31
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 6

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: RABGEF1
Fragment: 127-136
Site: 14-3-3_pSer132
Modification: (SEP)132
Sequence: INRQTSIETD
Construct: INRQT(pSer)IETD (crude synthetic peptide)
Average holdup BI: -0.02
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 113
P value: 0.24
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: RABGEF1
Fragment: 127-136
Site: 14-3-3_pSer132
Modification: (SEP)132
Sequence: INRQTSIETD
Construct: INRQT(pSer)IETD (crude synthetic peptide)
Average holdup BI: -0.03
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 142
P value: 0.49
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: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: RABGEF1
Fragment: 127-136
Site: 14-3-3_pSer132
Modification: (SEP)132
Sequence: INRQTSIETD
Construct: INRQT(pSer)IETD (crude synthetic peptide)
Average holdup BI: 0.03
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 111
P value: 0.42
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: RABGEF1
Fragment: 372-381
Site: 14-3-3_pSer377
Modification: (SEP)377
Sequence: QSLNLSQEDF
Construct: QSLNL(pSer)QEDF (crude synthetic peptide)
Average holdup BI: 0.09
Holdup BI standard deviation: 0.09
Immobilized partner concentration (10-6M): 113
P value: 1.11
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))

Experiment series 10

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: RABGEF1
Fragment: 372-381
Site: 14-3-3_pSer377
Modification: (SEP)377
Sequence: QSLNLSQEDF
Construct: QSLNL(pSer)QEDF (crude synthetic peptide)
Average holdup BI: -0.19
Holdup BI standard deviation: 0.09
Immobilized partner concentration (10-6M): 142
P value: 2.79
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: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: RABGEF1
Fragment: 372-381
Site: 14-3-3_pSer377
Modification: (SEP)377
Sequence: QSLNLSQEDF
Construct: QSLNL(pSer)QEDF (crude synthetic peptide)
Average holdup BI: -0.05
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 111
P value: 0.87
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 12

Protein A

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: RABGEF1
Fragment: 384-393
Site: 14-3-3_pThr389
Modification: (TPO)389
Sequence: YMSGQTSPRK
Construct: YMSGQ(pThr)SPRK (crude synthetic peptide)
Average holdup BI: 0.06
Holdup BI standard deviation: 0.64
Immobilized partner concentration (10-6M): 142
P value: 0.07
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))