Interaction Details


Details of experiments between O60292 and Q04917
Note that the results of all experiments are listed, regardless of the modification states of the fragments.


Experiment series 1

Protein A

Protein: SIPA1L3
Fragment: 1120-1129
Site: 14-3-3_pSer1125
Modification: (SEP)1125
Sequence: VPFRESQPLH
Construct: VPFRE(pSer)QPLH (crude synthetic peptide)

Protein B

Protein: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta
Average holdup BI: 0.04
Holdup BI standard deviation: 0.11
Immobilized partner concentration (10-6M): 130
P value: 0.37
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 2

Protein A

Protein: SIPA1L3
Fragment: 1130-1139
Site: 14-3-3_pSer1135
Modification: (SEP)1135
Sequence: SKRPVSFPET
Construct: SKRPV(pSer)FPET (crude synthetic peptide)

Protein B

Protein: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta
Average holdup BI: 0.63
Holdup BI standard deviation: 0.08
Immobilized partner concentration (10-6M): 130
P value: 5.73
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.12
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: SIPA1L3
Fragment: 1377-1386
Site: 14-3-3_pSer1382
Modification: (SEP)1382
Sequence: RPKLYSSGSS
Construct: biotin-ttds-RPKLY(pSer)SGSS (HPLC-purified synthetic peptide)

Protein B

Protein: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta
Measured dissociation constant (10-6M): 165
Standard deviation of dissociation constant (10-6M): 14
Experiment method: competitive Fluorescence Polarization
Details of affinity fitting: Competitive binding equation in ProFit
Date of measurement: 2022-01-19
Number of measurements: 3
Measured pKd: 3.78
DOI: 10.1101/2022.12.08.519103

Experiment series 4

Protein A

Protein: SIPA1L3
Fragment: 1377-1386
Site: 14-3-3_pSer1382
Modification: (SEP)1382
Sequence: RPKLYSSGSS
Construct: RPKLY(pSer)SGSS (crude synthetic peptide)

Protein B

Protein: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta
Average holdup BI: 0.29
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 130
P value: 4.51
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.5
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: SIPA1L3
Fragment: 1468-1477
Site: 14-3-3_pSer1473
Modification: (SEP)1473
Sequence: RPLPFSDPKK
Construct: RPLPF(pSer)DPKK (crude synthetic peptide)

Protein B

Protein: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta
Average holdup BI: 0.64
Holdup BI standard deviation: 0.08
Immobilized partner concentration (10-6M): 130
P value: 5.77
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.13
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: SIPA1L3
Fragment: 1539-1548
Site: 14-3-3_pSer1544
Modification: (SEP)1544
Sequence: LQRTLSDESL
Construct: LQRTL(pSer)DESL (crude synthetic peptide)

Protein B

Protein: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta
Average holdup BI: 0.1
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 130
P value: 3.5
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 7

Protein A

Protein: SIPA1L3
Fragment: 167-176
Site: 14-3-3_pSer172
Modification: (SEP)172
Sequence: RHRSSSEITL
Construct: RHRSS(pSer)EITL (crude synthetic peptide)

Protein B

Protein: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta
Average holdup BI: 0.49
Holdup BI standard deviation: 0.14
Immobilized partner concentration (10-6M): 130
P value: 3.41
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.87
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: SIPA1L3
Fragment: 386-395
Site: 14-3-3_pSer391
Modification: (SEP)391
Sequence: ASRAHSLGGL
Construct: ASRAH(pSer)LGGL (crude synthetic peptide)

Protein B

Protein: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta
Average holdup BI: 0.68
Holdup BI standard deviation: 0.1
Immobilized partner concentration (10-6M): 130
P value: 5.36
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.2
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 9

Protein A

Protein: SIPA1L3
Fragment: 92-101
Site: 14-3-3_pSer97
Modification: (SEP)97
Sequence: ALREHSNPSP
Construct: ALREH(pSer)NPSP (crude synthetic peptide)

Protein B

Protein: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta
Average holdup BI: -0.07
Holdup BI standard deviation: 0.09
Immobilized partner concentration (10-6M): 130
P value: 0.77
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))