Interaction Details


Details of experiments between Q9P0L2 and P31947
Note that the results of all experiments are listed, regardless of the modification states of the fragments.


Experiment series 1

Protein A

Protein: MARK1
Fragment: 644-653
Site: 14-3-3_pSer649
Modification: (SEP)649
Sequence: ARRGTSTGII
Construct: ARRGT(pSer)TGII (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.27
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 142
P value: 4.65
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.42
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: MARK1
Fragment: 644-653
Site: 14-3-3_pSer649
Modification: (SEP)649
Sequence: ARRGTSTGII
Construct: ARRGT(pSer)TGII (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.21
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 113
P value: 5.06
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: MARK1
Fragment: 210-219
Site: 14-3-3_pThr215
Modification: (TPO)215
Sequence: GNKLDTFCGS
Construct: GNKLD(pThr)FCGS (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Measured dissociation constant (10-6M): <100
Immobilized partner concentration (10-6M): 113
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))

Experiment series 4

Protein A

Protein: MARK1
Fragment: 60-69
Site: 14-3-3_pThr65
Modification: (TPO)65
Sequence: YRLQKTIGKG
Construct: YRLQK(pThr)IGKG (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.02
Holdup BI standard deviation: 0.09
Immobilized partner concentration (10-6M): 113
P value: 0.17
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: MARK1
Fragment: 60-69
Site: 14-3-3_pThr65
Modification: (TPO)65
Sequence: YRLQKTIGKG
Construct: YRLQK(pThr)IGKG (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.01
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 142
P value: 0.17
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: MARK1
Fragment: 644-653
Site: 14-3-3_pSer649
Modification: (SEP)649
Sequence: ARRGTSTGII
Construct: ARRGT(pSer)TGII (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.27
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 111
P value: 5.36
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.51
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: MARK1
Fragment: 60-69
Site: 14-3-3_pThr65
Modification: (TPO)65
Sequence: YRLQKTIGKG
Construct: YRLQK(pThr)IGKG (crude synthetic peptide)

Protein B

Protein: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma
Average holdup BI: 0.14
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 111
P value: 2.22
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))