Interaction Details


Details of experiments between Q9H6H4 and P61981
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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.92
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 129
P value: 3.65
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.98
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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.84
Holdup BI standard deviation: 0.11
Immobilized partner concentration (10-6M): 134
P value: 2.51
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.6
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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.93
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 129
P value: 9.57
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 5.01
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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.91
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 134
P value: 5.73
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.9
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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: -0.04
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 134
P value: 0.94
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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: -0.12
Holdup BI standard deviation: 0.17
Immobilized partner concentration (10-6M): 129
P value: 0.92
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: 245-254
Site: 14-3-3_pThr250
Modification: (TPO)250
Sequence: RTRKKTVPSD
Construct: RTRKK(pThr)VPSD (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: -0.27
Holdup BI standard deviation: 1.52
Immobilized partner concentration (10-6M): 129
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))

Experiment series 8

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: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.89
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 115
P value: 3.53
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.86
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: 219-228
Site: 14-3-3_pSer224
Modification: (SEP)224
Sequence: LIRSQSLRVV
Construct: LIRSQ(pSer)LRVV (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.91
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 115
P value: 6.97
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.95
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: REEP4
Fragment: 248-257
Site: 14-3-3_pSer253
Modification: (SEP)253
Sequence: KKTVPSDVDS
Construct: KKTVP(pSer)DVDS (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: -0.16
Holdup BI standard deviation: 0.17
Immobilized partner concentration (10-6M): 115
P value: 1.05
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))