Interaction Details


Details of experiments between Q96FS4 and P31946
Note that the results of all experiments are listed, regardless of the modification states of the fragments.


Experiment series 1

Protein A

Protein: SIPA1
Fragment: 50-59
Site: 14-3-3_pSer55
Modification: (SEP)55
Sequence: LLRSGSDAGE
Construct: LLRSG(pSer)DAGE (crude synthetic peptide)

Protein B

Protein: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 134
P value: 0.08
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 2

Protein A

Protein: SIPA1
Fragment: 50-59
Site: 14-3-3_pSer55
Modification: (SEP)55
Sequence: LLRSGSDAGE
Construct: LLRSG(pSer)DAGE (crude synthetic peptide)

Protein B

Protein: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: -0.14
Holdup BI standard deviation: 0.25
Immobilized partner concentration (10-6M): 102
P value: 0.73
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 3

Protein A

Protein: SIPA1
Fragment: 554-563
Site: 14-3-3_pSer559
Modification: (SEP)559
Sequence: RFGLPSLGGR
Construct: RFGLP(pSer)LGGR (crude synthetic peptide)

Protein B

Protein: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.23
Holdup BI standard deviation: 0.55
Immobilized partner concentration (10-6M): 134
P value: 0.34
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 4

Protein A

Protein: SIPA1
Fragment: 554-563
Site: 14-3-3_pSer559
Modification: (SEP)559
Sequence: RFGLPSLGGR
Construct: RFGLP(pSer)LGGR (crude synthetic peptide)

Protein B

Protein: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: -0.15
Holdup BI standard deviation: 0.31
Immobilized partner concentration (10-6M): 102
P value: 0.5
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 5

Protein A

Protein: SIPA1
Fragment: 675-684
Site: 14-3-3_pSer680
Modification: (SEP)680
Sequence: RLQLVSRGCE
Construct: RLQLV(pSer)RGCE (crude synthetic peptide)

Protein B

Protein: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: -0.01
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 134
P value: 0.04
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: SIPA1
Fragment: 832-841
Site: 14-3-3_pSer837
Modification: (SEP)837
Sequence: LHSQNSLSPR
Construct: LHSQN(pSer)LSPR (crude synthetic peptide)

Protein B

Protein: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.02
Holdup BI standard deviation: 0.14
Immobilized partner concentration (10-6M): 134
P value: 0.08
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: SIPA1
Fragment: 832-841
Site: 14-3-3_pSer837
Modification: (SEP)837
Sequence: LHSQNSLSPR
Construct: LHSQN(pSer)LSPR (crude synthetic peptide)

Protein B

Protein: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.05
Holdup BI standard deviation: 0.1
Immobilized partner concentration (10-6M): 102
P value: 0.43
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: SIPA1
Fragment: 903-912
Site: 14-3-3_pSer908
Modification: (SEP)908
Sequence: LPRTLSLRNS
Construct: LPRTL(pSer)LRNS (crude synthetic peptide)

Protein B

Protein: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.54
Holdup BI standard deviation: 0.08
Immobilized partner concentration (10-6M): 134
P value: 3.88
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.95
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: SIPA1
Fragment: 903-912
Site: 14-3-3_pSer908
Modification: (SEP)908
Sequence: LPRTLSLRNS
Construct: LPRTL(pSer)LRNS (crude synthetic peptide)

Protein B

Protein: YWHAB
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3beta
Average holdup BI: 0.47
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 102
P value: 5.62
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.94
DOI: 10.1101/2022.12.08.519103
Experimental Details

-log10(P Two-sided unpaired T-test (4 vs 5))