Interaction Details


Details of experiments between P31947 and Q7Z3C6
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: ATG9A
Fragment: 9-18
Site: 14-3-3_pSer14
Modification: (SEP)14
Sequence: QRLEASYSDS
Construct: QRLEA(pSer)YSDS (crude synthetic peptide)
Average holdup BI: -0.04
Holdup BI standard deviation: 0.08
Immobilized partner concentration (10-6M): 113
P value: 0.45
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: ATG9A
Fragment: 9-18
Site: 14-3-3_pSer14
Modification: (SEP)14
Sequence: QRLEASYSDS
Construct: QRLEA(pSer)YSDS (crude synthetic peptide)
Average holdup BI: -0.04
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 142
P value: 0.63
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: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: ATG9A
Fragment: 9-18
Site: 14-3-3_pSer14
Modification: (SEP)14
Sequence: QRLEASYSDS
Construct: QRLEA(pSer)YSDS (crude synthetic peptide)
Average holdup BI: -0.03
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 111
P value: 0.59
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: ATG9A
Fragment: 736-745
Site: 14-3-3_pSer741
Modification: (SEP)741
Sequence: DESGESAPDE
Construct: DESGE(pSer)APDE (crude synthetic peptide)
Average holdup BI: 0.65
Holdup BI standard deviation: 0.18
Immobilized partner concentration (10-6M): 113
P value: 3.27
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.22
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: ATG9A
Fragment: 736-745
Site: 14-3-3_pSer741
Modification: (SEP)741
Sequence: DESGESAPDE
Construct: DESGE(pSer)APDE (crude synthetic peptide)
Average holdup BI: 0.7
Holdup BI standard deviation: 0.14
Immobilized partner concentration (10-6M): 142
P value: 3.61
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.2
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: ATG9A
Fragment: 750-759
Site: 14-3-3_pSer755
Modification: (SEP)755
Sequence: ARAPQSIPRS
Construct: ARAPQ(pSer)IPRS (crude synthetic peptide)
Average holdup BI: 0.11
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 113
P value: 1.55
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: ATG9A
Fragment: 750-759
Site: 14-3-3_pSer755
Modification: (SEP)755
Sequence: ARAPQSIPRS
Construct: ARAPQ(pSer)IPRS (crude synthetic peptide)
Average holdup BI: 0.09
Holdup BI standard deviation: 0.08
Immobilized partner concentration (10-6M): 142
P value: 1.29
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: ATG9A
Fragment: 750-759
Site: 14-3-3_pSer755
Modification: (SEP)755
Sequence: ARAPQSIPRS
Construct: ARAPQ(pSer)IPRS (crude synthetic peptide)
Average holdup BI: 0.2
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 111
P value: 3.6
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: ATG9A
Fragment: 756-765
Site: 14-3-3_pSer761
Modification: (SEP)761
Sequence: IPRSASYPCA
Construct: IPRSA(pSer)YPCA (crude synthetic peptide)
Average holdup BI: -0.34
Holdup BI standard deviation: 0.64
Immobilized partner concentration (10-6M): 113
P value: 0.67
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: ATG9A
Fragment: 756-765
Site: 14-3-3_pSer761
Modification: (SEP)761
Sequence: IPRSASYPCA
Construct: IPRSA(pSer)YPCA (crude synthetic peptide)
Average holdup BI: 0.02
Holdup BI standard deviation: 0.13
Immobilized partner concentration (10-6M): 142
P value: 0.08
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
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: ATG9A
Fragment: 823-832
Site: 14-3-3_pSer828
Modification: (SEP)828
Sequence: VPEEGSEDEL
Construct: VPEEG(pSer)EDEL (crude synthetic peptide)
Average holdup BI: -0.19
Holdup BI standard deviation: 0.58
Immobilized partner concentration (10-6M): 113
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 12

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: ATG9A
Fragment: 823-832
Site: 14-3-3_pSer828
Modification: (SEP)828
Sequence: VPEEGSEDEL
Construct: VPEEG(pSer)EDEL (crude synthetic peptide)
Average holdup BI: 0.04
Holdup BI standard deviation: 0.92
Immobilized partner concentration (10-6M): 111
P value: 0.03
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))