Interaction Details


Details of experiments between Q04917 and Q8IZ21
Note that the results of all experiments are listed, regardless of the modification states of the fragments.


Experiment series 1

Protein A

Protein: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta

Protein B

Protein: PHACTR4
Fragment: 126-135
Site: 14-3-3_pSer131
Modification: (SEP)131
Sequence: PVRLASLRKA
Construct: PVRLA(pSer)LRKA (crude synthetic peptide)
Average holdup BI: 0.13
Holdup BI standard deviation: 0.12
Immobilized partner concentration (10-6M): 130
P value: 1.23
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: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta

Protein B

Protein: PHACTR4
Fragment: 142-151
Site: 14-3-3_pSer147
Modification: (SEP)147
Sequence: KKRLGSTGSQ
Construct: KKRLG(pSer)TGSQ (crude synthetic peptide)
Average holdup BI: -0.12
Holdup BI standard deviation: 0.35
Immobilized partner concentration (10-6M): 130
P value: 0.34
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: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta

Protein B

Protein: PHACTR4
Fragment: 265-274
Site: 14-3-3_pSer270
Modification: (SEP)270
Sequence: PAHRNSNPVI
Construct: PAHRN(pSer)NPVI (crude synthetic peptide)
Average holdup BI: -0.01
Holdup BI standard deviation: 0.13
Immobilized partner concentration (10-6M): 130
P value: 0.06
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 4

Protein A

Protein: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta

Protein B

Protein: PHACTR4
Fragment: 438-447
Site: 14-3-3_pSer443
Modification: (SEP)443
Sequence: SHRAHSLLFE
Construct: SHRAH(pSer)LLFE (crude synthetic peptide)
Average holdup BI: 0.07
Holdup BI standard deviation: 0.35
Immobilized partner concentration (10-6M): 130
P value: 0.14
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: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta

Protein B

Protein: PHACTR4
Fragment: 459-468
Site: 14-3-3_pSer464
Modification: (SEP)464
Sequence: LGRTRSLPIT
Construct: LGRTR(pSer)LPIT (crude synthetic peptide)
Average holdup BI: 0.98
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 130
P value: 6.97
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 5.52
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: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta

Protein B

Protein: PHACTR4
Fragment: 552-561
Site: 14-3-3_pSer557
Modification: (SEP)557
Sequence: LNHRPSEPEL
Construct: LNHRP(pSer)EPEL (crude synthetic peptide)
Average holdup BI: 0.04
Holdup BI standard deviation: 0.08
Immobilized partner concentration (10-6M): 130
P value: 0.44
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: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta

Protein B

Protein: PHACTR4
Fragment: 585-594
Site: 14-3-3_pSer590
Modification: (SEP)590
Sequence: LIRRLSQRPT
Construct: LIRRL(pSer)QRPT (crude synthetic peptide)
Average holdup BI: 0
Holdup BI standard deviation: 0.08
Immobilized partner concentration (10-6M): 130
P value: 0.03
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: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta

Protein B

Protein: PHACTR4
Fragment: 623-632
Site: 14-3-3_pSer628
Modification: (SEP)628
Sequence: LTRKLSQRPT
Construct: LTRKL(pSer)QRPT (crude synthetic peptide)
Average holdup BI: 0.02
Holdup BI standard deviation: 0.24
Immobilized partner concentration (10-6M): 130
P value: 0.08
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 9

Protein A

Protein: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta

Protein B

Protein: PHACTR4
Fragment: 65-74
Site: 14-3-3_pSer70
Modification: (SEP)70
Sequence: LERKISMRKP
Construct: LERKI(pSer)MRKP (crude synthetic peptide)
Average holdup BI: 0.1
Holdup BI standard deviation: 0.14
Immobilized partner concentration (10-6M): 130
P value: 0.64
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))