Interaction Details


Details of experiments between Q9UJF2 and Q04917
Note that the results of all experiments are listed, regardless of the modification states of the fragments.


Experiment series 1

Protein A

Protein: RASAL2
Fragment: 129-138
Site: 14-3-3_pSer134
Modification: (SEP)134
Sequence: LRRTVSVPSE
Construct: LRRTV(pSer)VPSE (crude synthetic peptide)

Protein B

Protein: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta
Average holdup BI: 0.84
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 130
P value: 6.82
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.62
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: RASAL2
Fragment: 731-740
Site: 14-3-3_pSer736
Modification: (SEP)736
Sequence: IRLTGSQLSI
Construct: IRLTG(pSer)QLSI (crude synthetic peptide)

Protein B

Protein: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta
Average holdup BI: -0.03
Holdup BI standard deviation: 0.65
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 3

Protein A

Protein: RASAL2
Fragment: 734-743
Site: 14-3-3_pSer739
Modification: (SEP)739
Sequence: TGSQLSITQV
Construct: TGSQL(pSer)ITQV (crude synthetic peptide)

Protein B

Protein: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta
Average holdup BI: -0.06
Holdup BI standard deviation: 0.11
Immobilized partner concentration (10-6M): 130
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 4

Protein A

Protein: RASAL2
Fragment: 836-845
Site: 14-3-3_pSer841
Modification: (SEP)841
Sequence: KRSTQSEDFS
Construct: KRSTQ(pSer)EDFS (crude synthetic peptide)

Protein B

Protein: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta
Average holdup BI: 0.48
Holdup BI standard deviation: 0.09
Immobilized partner concentration (10-6M): 130
P value: 3.57
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.85
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: RASAL2
Fragment: 882-891
Site: 14-3-3_pSer887
Modification: (SEP)887
Sequence: AKAPPSLPHS
Construct: AKAPP(pSer)LPHS (crude synthetic peptide)

Protein B

Protein: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta
Average holdup BI: 0.11
Holdup BI standard deviation: 0.17
Immobilized partner concentration (10-6M): 130
P value: 0.62
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: RASAL2
Fragment: 84-93
Site: 14-3-3_pSer89
Modification: (SEP)89
Sequence: LKESRSHESL
Construct: LKESR(pSer)HESL (crude synthetic peptide)

Protein B

Protein: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta
Average holdup BI: 0.25
Holdup BI standard deviation: 0.1
Immobilized partner concentration (10-6M): 130
P value: 2.75
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 7

Protein A

Protein: RASAL2
Fragment: 612-621
Site: 14-3-3_pThr617
Modification: (TPO)617
Sequence: ITKSLTNPTP
Construct: ITKSL(pThr)NPTP (crude synthetic peptide)

Protein B

Protein: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta
Average holdup BI: 0.77
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 130
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: 4.4
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: RASAL2
Fragment: 844-853
Site: 14-3-3_pThr849
Modification: (TPO)849
Sequence: FSRRHTVPDR
Construct: FSRRH(pThr)VPDR (crude synthetic peptide)

Protein B

Protein: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta
Average holdup BI: 0.87
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 130
P value: 3.9
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.72
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: RASAL2
Fragment: 936-945
Site: 14-3-3_pThr941
Modification: (TPO)941
Sequence: IQRQQTQQVQ
Construct: IQRQQ(pThr)QQVQ (crude synthetic peptide)

Protein B

Protein: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta
Average holdup BI: 0.05
Holdup BI standard deviation: 0.22
Immobilized partner concentration (10-6M): 130
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))