Interaction Details


Details of experiments between Q7Z401 and P27348
Note that the results of all experiments are listed, regardless of the modification states of the fragments.


Experiment series 1

Protein A

Protein: DENND4A
Fragment: 1030-1039
Site: 14-3-3_pSer1035
Modification: (SEP)1035
Sequence: RNRNLSGGVL
Construct: RNRNL(pSer)GGVL (crude synthetic peptide)

Protein B

Protein: YWHAQ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3tau
Average holdup BI: 0.24
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 130
P value: 3.48
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.4
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: DENND4A
Fragment: 1146-1155
Site: 14-3-3_pSer1151
Modification: (SEP)1151
Sequence: IQRMNSSFSV
Construct: IQRMN(pSer)SFSV (crude synthetic peptide)

Protein B

Protein: YWHAQ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3tau
Average holdup BI: 0.08
Holdup BI standard deviation: 0.08
Immobilized partner concentration (10-6M): 130
P value: 1.03
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 3

Protein A

Protein: DENND4A
Fragment: 1220-1229
Site: 14-3-3_pSer1225
Modification: (SEP)1225
Sequence: TSRTPSIDLQ
Construct: TSRTP(pSer)IDLQ (crude synthetic peptide)

Protein B

Protein: YWHAQ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3tau
Average holdup BI: -0.01
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 130
P value: 0.11
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: DENND4A
Fragment: 1276-1285
Site: 14-3-3_pSer1281
Modification: (SEP)1281
Sequence: RDRLWSSPAF
Construct: RDRLW(pSer)SPAF (crude synthetic peptide)

Protein B

Protein: YWHAQ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3tau
Average holdup BI: 0.88
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 130
P value: 3.67
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.76
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: DENND4A
Fragment: 1503-1512
Site: 14-3-3_pSer1508
Modification: (SEP)1508
Sequence: RYFLKSSPST
Construct: RYFLK(pSer)SPST (crude synthetic peptide)

Protein B

Protein: YWHAQ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3tau
Average holdup BI: 0.68
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 130
P value: 7.03
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.21
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: DENND4A
Fragment: 1586-1595
Site: 14-3-3_pSer1591
Modification: (SEP)1591
Sequence: RSISTSGPLD
Construct: RSIST(pSer)GPLD (crude synthetic peptide)

Protein B

Protein: YWHAQ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3tau
Average holdup BI: 0.16
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 130
P value: 4.71
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: DENND4A
Fragment: 1585-1594
Site: 14-3-3_pThr1590
Modification: (TPO)1590
Sequence: ARSISTSGPL
Construct: ARSIS(pThr)SGPL (crude synthetic peptide)

Protein B

Protein: YWHAQ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3tau
Average holdup BI: 0.42
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 130
P value: 6.66
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.74
DOI: 10.1101/2022.12.08.519103
Experimental Details

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