Interaction Details


Details of experiments between Q9H6H4 and P62258
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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.72
Holdup BI standard deviation: 0.09
Immobilized partner concentration (10-6M): 143
P value: 2.97
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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.52
Holdup BI standard deviation: 0.22
Immobilized partner concentration (10-6M): 116
P value: 1.48
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.97
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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.65
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 116
P value: 4.81
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.2
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: REEP4
Fragment: 219-228
Site: 14-3-3_pSer224
Modification: (SEP)224
Sequence: LIRSQSLRVV
Construct: LIRSQ(pSer)LRVV (crude synthetic peptide)

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.66
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 143
P value: 7.94
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.14
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: REEP4
Fragment: 248-257
Site: 14-3-3_pSer253
Modification: (SEP)253
Sequence: KKTVPSDVDS
Construct: KKTVP(pSer)DVDS (crude synthetic peptide)

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.11
Holdup BI standard deviation: 0.09
Immobilized partner concentration (10-6M): 116
P value: 1.3
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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: -0.04
Holdup BI standard deviation: 0.19
Immobilized partner concentration (10-6M): 143
P value: 0.16
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))