Interaction Details


Details of experiments between Q9Y2U5 and P63104
Note that the results of all experiments are listed, regardless of the modification states of the fragments.


Experiment series 1

Protein A

Protein: MAP3K2
Fragment: 148-157
Site: 14-3-3_pSer153
Modification: (SEP)153
Sequence: RKKRLSIIGP
Construct: RKKRL(pSer)IIGP (crude synthetic peptide)

Protein B

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta
Average holdup BI: 0.22
Holdup BI standard deviation: 0.35
Immobilized partner concentration (10-6M): 132
P value: 0.58
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: MAP3K2
Fragment: 234-243
Site: 14-3-3_pSer239
Modification: (SEP)239
Sequence: MPRAQSYPDN
Construct: MPRAQ(pSer)YPDN (crude synthetic peptide)

Protein B

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta
Average holdup BI: 0.75
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 132
P value: 12.68
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.36
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: MAP3K2
Fragment: 243-252
Site: 14-3-3_pSer248
Modification: (SEP)248
Sequence: NHQEFSDYDN
Construct: NHQEF(pSer)DYDN (crude synthetic peptide)

Protein B

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta
Average holdup BI: -0.11
Holdup BI standard deviation: 0.5
Immobilized partner concentration (10-6M): 132
P value: 0.18
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: MAP3K2
Fragment: 488-497
Site: 14-3-3_pSer493
Modification: (SEP)493
Sequence: NILRDSTGNV
Construct: NILRD(pSer)TGNV (crude synthetic peptide)

Protein B

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta
Average holdup BI: 0.04
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 132
P value: 1.81
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: MAP3K2
Fragment: 515-524
Site: 14-3-3_pSer520
Modification: (SEP)520
Sequence: GTGMKSVTGT
Construct: GTGMK(pSer)VTGT (crude synthetic peptide)

Protein B

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta
Average holdup BI: 0.03
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 132
P value: 0.51
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 6

Protein A

Protein: MAP3K2
Fragment: 258-267
Site: 14-3-3_pThr263
Modification: (TPO)263
Sequence: FGKGGTYPRR
Construct: FGKGG(pThr)YPRR (crude synthetic peptide)

Protein B

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta
Average holdup BI: 0.87
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 132
P value: 7.36
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.71
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: MAP3K2
Fragment: 278-287
Site: 14-3-3_pThr283
Modification: (TPO)283
Sequence: NDGRKTFPRA
Construct: NDGRK(pThr)FPRA (crude synthetic peptide)

Protein B

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta
Average holdup BI: 0.69
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 132
P value: 8.84
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.22
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: MAP3K2
Fragment: 517-526
Site: 14-3-3_pThr522
Modification: (TPO)522
Sequence: GMKSVTGTPY
Construct: GMKSV(pThr)GTPY (crude synthetic peptide)

Protein B

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta
Average holdup BI: 0.53
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 132
P value: 6.02
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.93
DOI: 10.1101/2022.12.08.519103
Experimental Details

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