Interaction Details


Details of experiments between Q9UJF2 and P61981
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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.92
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 129
P value: 7.94
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.95
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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.9
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 134
P value: 9.6
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.84
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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.06
Holdup BI standard deviation: 0.39
Immobilized partner concentration (10-6M): 134
P value: 0.12
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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: -0.02
Holdup BI standard deviation: 0.6
Immobilized partner concentration (10-6M): 129
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 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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.07
Holdup BI standard deviation: 0.08
Immobilized partner concentration (10-6M): 134
P value: 0.87
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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.01
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 129
P value: 0.14
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: 836-845
Site: 14-3-3_pSer841
Modification: (SEP)841
Sequence: KRSTQSEDFS
Construct: KRSTQ(pSer)EDFS (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.58
Holdup BI standard deviation: 0.09
Immobilized partner concentration (10-6M): 129
P value: 2.83
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.03
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: 882-891
Site: 14-3-3_pSer887
Modification: (SEP)887
Sequence: AKAPPSLPHS
Construct: AKAPP(pSer)LPHS (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.19
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 134
P value: 3.01
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 9

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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.16
Holdup BI standard deviation: 0.17
Immobilized partner concentration (10-6M): 129
P value: 0.92
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: RASAL2
Fragment: 84-93
Site: 14-3-3_pSer89
Modification: (SEP)89
Sequence: LKESRSHESL
Construct: LKESR(pSer)HESL (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.36
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 129
P value: 4.55
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.63
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: 84-93
Site: 14-3-3_pSer89
Modification: (SEP)89
Sequence: LKESRSHESL
Construct: LKESR(pSer)HESL (crude synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.31
Holdup BI standard deviation: 0.16
Immobilized partner concentration (10-6M): 134
P value: 1.75
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 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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.83
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 134
P value: 7.96
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.57
DOI: 10.1101/2022.12.08.519103
Experimental Details

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


Experiment series 13

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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.82
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 129
P value: 8.3
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.56
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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.94
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 129
P value: 4.1
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 5.06
DOI: 10.1101/2022.12.08.519103
Experimental Details

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


Experiment series 15

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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.92
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 134
P value: 7.82
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.95
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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.07
Holdup BI standard deviation: 0.17
Immobilized partner concentration (10-6M): 134
P value: 0.34
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 17

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: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.16
Holdup BI standard deviation: 0.22
Immobilized partner concentration (10-6M): 129
P value: 0.69
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 18

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: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.94
Holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 115
P value: 7.6
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 5.14
DOI: 10.1101/2022.12.08.519103
Experimental Details

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


Experiment series 19

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: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: -0.02
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 115
P value: 0.36
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 20

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: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.14
Holdup BI standard deviation: 0.1
Immobilized partner concentration (10-6M): 115
P value: 1.34
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 21

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: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: -0.01
Holdup BI standard deviation: 0.1
Immobilized partner concentration (10-6M): 115
P value: 0.06
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 22

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: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.19
Holdup BI standard deviation: 0.09
Immobilized partner concentration (10-6M): 115
P value: 2.21
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 23

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: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.24
Holdup BI standard deviation: 0.2
Immobilized partner concentration (10-6M): 115
P value: 1.22
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 24

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: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.83
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 115
P value: 8.55
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.63
DOI: 10.1101/2022.12.08.519103
Experimental Details

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


Experiment series 25

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: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.84
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 115
P value: 3.72
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.67
DOI: 10.1101/2022.12.08.519103
Experimental Details

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


Experiment series 26

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: YWHAG
Fragment: 1-247
Site: 14-3-3
Modification: 100_uM_FCA
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Average holdup BI: 0.08
Holdup BI standard deviation: 0.38
Immobilized partner concentration (10-6M): 115
P value: 0.16
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))