Interaction Details


Details of experiments between Q9UJF2 and P62258
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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.67
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 143
P value: 6.65
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.16
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: 129-138
Site: 14-3-3_pSer134
Modification: (SEP)134
Sequence: LRRTVSVPSE
Construct: LRRTV(pSer)VPSE (crude synthetic peptide)

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.62
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 116
P value: 7.53
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.15
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 3

Protein A

Protein: RASAL2
Fragment: 695-704
Site: 14-3-3_pSer700
Modification: (SEP)700
Sequence: DERESSLPNG
Construct: DERES(pSer)LPNG (crude synthetic peptide)

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: -0.19
Holdup BI standard deviation: 0.16
Immobilized partner concentration (10-6M): 116
P value: 1.46
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
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 4))

Experiment series 4

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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: -0.49
Holdup BI standard deviation: 0.3
Immobilized partner concentration (10-6M): 116
P value: 1.76
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
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 4))

Experiment series 5

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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: -0.04
Holdup BI standard deviation: 0.09
Immobilized partner concentration (10-6M): 116
P value: 0.38
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
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 4))

Experiment series 6

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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.01
Holdup BI standard deviation: 0.08
Immobilized partner concentration (10-6M): 143
P value: 0.05
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: RASAL2
Fragment: 882-891
Site: 14-3-3_pSer887
Modification: (SEP)887
Sequence: AKAPPSLPHS
Construct: AKAPP(pSer)LPHS (crude synthetic peptide)

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.03
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 116
P value: 0.55
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
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 4))

Experiment series 8

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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0
Holdup BI standard deviation: 0.21
Immobilized partner concentration (10-6M): 143
P value: 0.02
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: RASAL2
Fragment: 84-93
Site: 14-3-3_pSer89
Modification: (SEP)89
Sequence: LKESRSHESL
Construct: LKESR(pSer)HESL (crude synthetic peptide)

Protein B

Protein: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.06
Holdup BI standard deviation: 0.15
Immobilized partner concentration (10-6M): 116
P value: 0.35
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
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 4))

Experiment series 10

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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.03
Holdup BI standard deviation: 0.19
Immobilized partner concentration (10-6M): 143
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 11

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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.59
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 116
P value: 6.83
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.09
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 12

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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.63
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 143
P value: 6.8
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.08
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 13

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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.85
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 143
P value: 3.81
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.59
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 14

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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.8
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 116
P value: 7.51
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.55
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 15

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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: 0.1
Holdup BI standard deviation: 0.12
Immobilized partner concentration (10-6M): 116
P value: 0.69
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
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 4))

Experiment series 16

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: YWHAE
Fragment: 1-255
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3epsilon
Average holdup BI: -0.03
Holdup BI standard deviation: 0.23
Immobilized partner concentration (10-6M): 143
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))