Interaction Details


Details of experiments between P63104 and Q6WCQ1
Note that the results of all experiments are listed, regardless of the modification states of the fragments.


Experiment series 1

Protein A

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta

Protein B

Protein: MPRIP
Fragment: 1011-1020
Site: 14-3-3_pSer1016
Modification: (SEP)1016
Sequence: NIRSKSVIEQ
Construct: NIRSK(pSer)VIEQ (crude synthetic peptide)
Average holdup BI: 0.49
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 132
P value: 7.13
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.86
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: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta

Protein B

Protein: MPRIP
Fragment: 1011-1020
Site: 14-3-3_pSer1016
Modification: (SEP)1016
Sequence: NIRSKSLKEG
Construct: NIRSK(pSer)LKEG (crude synthetic peptide)
Average holdup BI: -0.04
Holdup BI standard deviation: 0.69
Immobilized partner concentration (10-6M): 132
P value: 0.04
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: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta

Protein B

Protein: MPRIP
Fragment: 264-273
Site: 14-3-3_pSer269
Modification: (SEP)269
Sequence: ESGYFSLEKT
Construct: ESGYF(pSer)LEKT (crude synthetic peptide)
Average holdup BI: 0.14
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 132
P value: 2.27
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: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta

Protein B

Protein: MPRIP
Fragment: 289-298
Site: 14-3-3_pSer294
Modification: (SEP)294
Sequence: SPPSPSTPNH
Construct: SPPSP(pSer)TPNH (crude synthetic peptide)
Average holdup BI: 0.15
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 132
P value: 2.82
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: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta

Protein B

Protein: MPRIP
Fragment: 296-305
Site: 14-3-3_pSer301
Modification: (SEP)301
Sequence: PNHRRSQVIE
Construct: PNHRR(pSer)QVIE (crude synthetic peptide)
Average holdup BI: -0.26
Holdup BI standard deviation: 0.72
Immobilized partner concentration (10-6M): 132
P value: 0.2
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: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta

Protein B

Protein: MPRIP
Fragment: 988-997
Site: 14-3-3_pSer993
Modification: (SEP)993
Sequence: IMKSKSNPDF
Construct: IMKSK(pSer)NPDF (crude synthetic peptide)
Average holdup BI: 0.99
Holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 132
P value: 8.38
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 6.06
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: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta

Protein B

Protein: MPRIP
Fragment: 373-382
Site: 14-3-3_pThr378
Modification: (TPO)378
Sequence: LDRRSTEPSV
Construct: LDRRS(pThr)EPSV (crude synthetic peptide)
Average holdup BI: 0.63
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 132
P value: 8.28
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.11
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: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta

Protein B

Protein: MPRIP
Fragment: 537-546
Site: 14-3-3_pThr542
Modification: (TPO)542
Sequence: EGRSKTFDWA
Construct: EGRSK(pThr)FDWA (crude synthetic peptide)
Average holdup BI: 0.63
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 132
P value: 6.26
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.11
DOI: 10.1101/2022.12.08.519103
Experimental Details

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