Interaction Details


Details of experiments between P31947 and Q9NSK0
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: KLC4
Fragment: 549-558
Site: 14-3-3_pSer554
Modification: (SEP)554
Sequence: LQRSGSLGKI
Construct: LQRSG(pSer)LGKI (crude synthetic peptide)
Average holdup BI: 0.3
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 142
P value: 5.14
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.48
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: KLC4
Fragment: 549-558
Site: 14-3-3_pSer554
Modification: (SEP)554
Sequence: LQRSGSLGKI
Construct: LQRSG(pSer)LGKI (crude synthetic peptide)
Average holdup BI: 0.24
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 113
P value: 3.23
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.43
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: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: KLC4
Fragment: 549-558
Site: 14-3-3_pSer554
Modification: (SEP)554
Sequence: LQRSGSLGKI
Construct: LQRSG(pSer)LGKI (crude synthetic peptide)
Average holdup BI: 0.25
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 111
P value: 5.8
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.48
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: KLC4
Fragment: 561-570
Site: 14-3-3_pSer566
Modification: (SEP)566
Sequence: VLRRSSELLV
Construct: VLRRS(pSer)ELLV (crude synthetic peptide)
Average holdup BI: -0.14
Holdup BI standard deviation: 0.13
Immobilized partner concentration (10-6M): 113
P value: 1.21
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: SFN
Fragment: 1-248
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: KLC4
Fragment: 561-570
Site: 14-3-3_pSer566
Modification: (SEP)566
Sequence: VLRRSSELLV
Construct: VLRRS(pSer)ELLV (crude synthetic peptide)
Average holdup BI: -0.02
Holdup BI standard deviation: 0.09
Immobilized partner concentration (10-6M): 142
P value: 0.21
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: SFN
Fragment: 1-248
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: KLC4
Fragment: 561-570
Site: 14-3-3_pSer566
Modification: (SEP)566
Sequence: VLRRSSELLV
Construct: VLRRS(pSer)ELLV (crude synthetic peptide)
Average holdup BI: -0.02
Holdup BI standard deviation: 0.12
Immobilized partner concentration (10-6M): 111
P value: 0.15
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: KLC4
Fragment: 585-594
Site: 14-3-3_pSer590
Modification: (SEP)590
Sequence: MKRAASLNYL
Construct: MKRAA(pSer)LNYL (crude synthetic peptide)
Average holdup BI: 0.78
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 142
P value: 7
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.4
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
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: KLC4
Fragment: 585-594
Site: 14-3-3_pSer590
Modification: (SEP)590
Sequence: MKRAASLNYL
Construct: MKRAA(pSer)LNYL (crude synthetic peptide)
Average holdup BI: 0.72
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 113
P value: 5.05
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.36
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
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3sigma

Protein B

Protein: KLC4
Fragment: 585-594
Site: 14-3-3_pSer590
Modification: (SEP)590
Sequence: MKRAASLNYL
Construct: MKRAA(pSer)LNYL (crude synthetic peptide)
Average holdup BI: 0.71
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 111
P value: 6.03
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.35
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 4))