Interaction Details


Details of experiments between Q5T5U3 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: ARHGAP21
Fragment: 5-14
Site: 14-3-3_pSer10
Modification: (SEP)10
Sequence: RRTGLSEGDG
Construct: RRTGL(pSer)EGDG (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: 0.17
Holdup BI standard deviation: 1.24
Immobilized partner concentration (10-6M): 132
P value: 0.1
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: ARHGAP21
Fragment: 1487-1496
Site: 14-3-3_pSer1492
Modification: (SEP)1492
Sequence: KDEKISLGKE
Construct: KDEKI(pSer)LGKE (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: 0.07
Immobilized partner concentration (10-6M): 132
P value: 0.39
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: ARHGAP21
Fragment: 1492-1501
Site: 14-3-3_pSer1497
Modification: (SEP)1497
Sequence: SLGKESTPSE
Construct: SLGKE(pSer)TPSE (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: -0.02
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 132
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 4

Protein A

Protein: ARHGAP21
Fragment: 1737-1746
Site: 14-3-3_pSer1742
Modification: (SEP)1742
Sequence: MKKGKSTGSL
Construct: MKKGK(pSer)TGSL (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: 0.11
Immobilized partner concentration (10-6M): 132
P value: 0.32
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: ARHGAP21
Fragment: 228-237
Site: 14-3-3_pSer233
Modification: (SEP)233
Sequence: SKQQTSTPVL
Construct: SKQQT(pSer)TPVL (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: 0.27
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 132
P value: 8.24
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.44
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: ARHGAP21
Fragment: 454-463
Site: 14-3-3_pSer459
Modification: (SEP)459
Sequence: RQRSVSQERL
Construct: RQRSV(pSer)QERL (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: 0.21
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 132
P value: 3.72
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: ARHGAP21
Fragment: 52-61
Site: 14-3-3_pSer57
Modification: (SEP)57
Sequence: TLKRTSQGFG
Construct: TLKRT(pSer)QGFG (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: 0.05
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 132
P value: 0.81
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 8

Protein A

Protein: ARHGAP21
Fragment: 607-616
Site: 14-3-3_pSer612
Modification: (SEP)612
Sequence: LPRGISQDRS
Construct: LPRGI(pSer)QDRS (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: 0.67
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: ARHGAP21
Fragment: 666-675
Site: 14-3-3_pSer671
Modification: (SEP)671
Sequence: WVRTDSAPDQ
Construct: WVRTD(pSer)APDQ (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: 0.05
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 132
P value: 1.54
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 10

Protein A

Protein: ARHGAP21
Fragment: 1798-1807
Site: 14-3-3_pThr1803
Modification: (TPO)1803
Sequence: IRRRHTLGGH
Construct: IRRRH(pThr)LGGH (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: 0.25
Immobilized partner concentration (10-6M): 132
P value: 0.22
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 11

Protein A

Protein: ARHGAP21
Fragment: 975-984
Site: 14-3-3_pThr980
Modification: (TPO)980
Sequence: DKREQTTPSE
Construct: DKREQ(pThr)TPSE (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: -0.24
Holdup BI standard deviation: 1.2
Immobilized partner concentration (10-6M): 132
P value: 0.12
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))