Interaction Details


Details of experiments between Q9H6H4 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: REEP4
Fragment: 147-156
Site: 14-3-3_pSer152
Modification: (SEP)152
Sequence: RLRSFSMQDL
Construct: RLRSF(pSer)MQDL (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: 0.72
Holdup BI standard deviation: 0.1
Immobilized partner concentration (10-6M): 142
P value: 2.95
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.26
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: REEP4
Fragment: 147-156
Site: 14-3-3_pSer152
Modification: (SEP)152
Sequence: RLRSFSMQDL
Construct: RLRSF(pSer)MQDL (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: 0.66
Holdup BI standard deviation: 0.16
Immobilized partner concentration (10-6M): 113
P value: 1.96
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.23
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: REEP4
Fragment: 219-228
Site: 14-3-3_pSer224
Modification: (SEP)224
Sequence: LIRSQSLRVV
Construct: LIRSQ(pSer)LRVV (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: 0.83
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 142
P value: 8.52
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.52
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: REEP4
Fragment: 219-228
Site: 14-3-3_pSer224
Modification: (SEP)224
Sequence: LIRSQSLRVV
Construct: LIRSQ(pSer)LRVV (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: 0.78
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 113
P value: 5.33
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.51
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: REEP4
Fragment: 248-257
Site: 14-3-3_pSer253
Modification: (SEP)253
Sequence: KKTVPSDVDS
Construct: KKTVP(pSer)DVDS (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: 0.04
Holdup BI standard deviation: 0.03
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 6

Protein A

Protein: REEP4
Fragment: 248-257
Site: 14-3-3_pSer253
Modification: (SEP)253
Sequence: KKTVPSDVDS
Construct: KKTVP(pSer)DVDS (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: -0.07
Holdup BI standard deviation: 0.17
Immobilized partner concentration (10-6M): 142
P value: 0.45
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 7

Protein A

Protein: REEP4
Fragment: 147-156
Site: 14-3-3_pSer152
Modification: (SEP)152
Sequence: RLRSFSMQDL
Construct: RLRSF(pSer)MQDL (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: 0.73
Holdup BI standard deviation: 0.08
Immobilized partner concentration (10-6M): 111
P value: 3.05
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.37
DOI: 10.1101/2022.12.08.519103
Experimental Details

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


Experiment series 8

Protein A

Protein: REEP4
Fragment: 219-228
Site: 14-3-3_pSer224
Modification: (SEP)224
Sequence: LIRSQSLRVV
Construct: LIRSQ(pSer)LRVV (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: 0.79
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 111
P value: 6.5
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.53
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: REEP4
Fragment: 248-257
Site: 14-3-3_pSer253
Modification: (SEP)253
Sequence: KKTVPSDVDS
Construct: KKTVP(pSer)DVDS (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: 0.18
Immobilized partner concentration (10-6M): 111
P value: 0.81
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))