Interaction Details


Details of experiments between O75122 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: CLASP2
Fragment: 1007-1016
Site: 14-3-3_pSer1012
Modification: (SEP)1012
Sequence: SMPTHSSPRS
Construct: SMPTH(pSer)SPRS (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: 0.32
Holdup BI standard deviation: 0.22
Immobilized partner concentration (10-6M): 134
P value: 1.03
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 2

Protein A

Protein: CLASP2
Fragment: 1007-1016
Site: 14-3-3_pSer1012
Modification: (SEP)1012
Sequence: SMPTHSSPRS
Construct: SMPTH(pSer)SPRS (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: 0.24
Holdup BI standard deviation: 0.18
Immobilized partner concentration (10-6M): 129
P value: 1.42
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: CLASP2
Fragment: 1029-1038
Site: 14-3-3_pSer1034
Modification: (SEP)1034
Sequence: SPFNKSALKE
Construct: SPFNK(pSer)ALKE (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
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 134
P value: 0.03
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: CLASP2
Fragment: 1029-1038
Site: 14-3-3_pSer1034
Modification: (SEP)1034
Sequence: SPFNKSALKE
Construct: SPFNK(pSer)ALKE (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
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 129
P value: 0.01
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: CLASP2
Fragment: 1048-1057
Site: 14-3-3_pSer1053
Modification: (SEP)1053
Sequence: FPDDLSLDHS
Construct: FPDDL(pSer)LDHS (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: -0.03
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 134
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: CLASP2
Fragment: 1048-1057
Site: 14-3-3_pSer1053
Modification: (SEP)1053
Sequence: FPDDLSLDHS
Construct: FPDDL(pSer)LDHS (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: -0.09
Holdup BI standard deviation: 0.08
Immobilized partner concentration (10-6M): 129
P value: 1.2
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 7

Protein A

Protein: CLASP2
Fragment: 347-356
Site: 14-3-3_pSer352
Modification: (SEP)352
Sequence: VAGRVSAGSS
Construct: VAGRV(pSer)AGSS (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: -0.11
Holdup BI standard deviation: 0.23
Immobilized partner concentration (10-6M): 134
P value: 0.42
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: CLASP2
Fragment: 347-356
Site: 14-3-3_pSer352
Modification: (SEP)352
Sequence: VAGRVSAGSS
Construct: VAGRV(pSer)AGSS (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: 0.2
Immobilized partner concentration (10-6M): 129
P value: 0.29
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: CLASP2
Fragment: 355-364
Site: 14-3-3_pSer360
Modification: (SEP)360
Sequence: SSKASSLPGS
Construct: SSKAS(pSer)LPGS (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: 0.02
Immobilized partner concentration (10-6M): 129
P value: 9.49
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.55
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: CLASP2
Fragment: 355-364
Site: 14-3-3_pSer360
Modification: (SEP)360
Sequence: SSKASSLPGS
Construct: SSKAS(pSer)LPGS (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: 0.78
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 134
P value: 8.45
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.43
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 11

Protein A

Protein: CLASP2
Fragment: 359-368
Site: 14-3-3_pSer364
Modification: (SEP)364
Sequence: SSLPGSLQRS
Construct: SSLPG(pSer)LQRS (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: 0.03
Immobilized partner concentration (10-6M): 134
P value: 2.05
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: CLASP2
Fragment: 359-368
Site: 14-3-3_pSer364
Modification: (SEP)364
Sequence: SSLPGSLQRS
Construct: SSLPG(pSer)LQRS (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: 0.09
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 129
P value: 2.74
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 13

Protein A

Protein: CLASP2
Fragment: 365-374
Site: 14-3-3_pSer370
Modification: (SEP)370
Sequence: LQRSRSDIDV
Construct: LQRSR(pSer)DIDV (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: 0.52
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 129
P value: 6.43
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.92
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: CLASP2
Fragment: 365-374
Site: 14-3-3_pSer370
Modification: (SEP)370
Sequence: LQRSRSDIDV
Construct: LQRSR(pSer)DIDV (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: 0.14
Holdup BI standard deviation: 0.58
Immobilized partner concentration (10-6M): 134
P value: 0.18
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 15

Protein A

Protein: CLASP2
Fragment: 425-434
Site: 14-3-3_pSer430
Modification: (SEP)430
Sequence: VRAKLSAPLA
Construct: VRAKL(pSer)APLA (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: 0.46
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 129
P value: 5.48
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.81
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 16

Protein A

Protein: CLASP2
Fragment: 425-434
Site: 14-3-3_pSer430
Modification: (SEP)430
Sequence: VRAKLSAPLA
Construct: VRAKL(pSer)APLA (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: 0.42
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 134
P value: 5.19
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.74
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: CLASP2
Fragment: 450-459
Site: 14-3-3_pSer455
Modification: (SEP)455
Sequence: KMVSQSQPGS
Construct: KMVSQ(pSer)QPGS (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: 0.03
Immobilized partner concentration (10-6M): 134
P value: 5.8
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.53
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 18

Protein A

Protein: CLASP2
Fragment: 450-459
Site: 14-3-3_pSer455
Modification: (SEP)455
Sequence: KMVSQSQPGS
Construct: KMVSQ(pSer)QPGS (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: 0.8
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 129
P value: 9.51
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.48
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 19

Protein A

Protein: CLASP2
Fragment: 484-493
Site: 14-3-3_pSer489
Modification: (SEP)489
Sequence: LVNSASAQKR
Construct: LVNSA(pSer)AQKR (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: 0.18
Immobilized partner concentration (10-6M): 134
P value: 0.89
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: CLASP2
Fragment: 484-493
Site: 14-3-3_pSer489
Modification: (SEP)489
Sequence: LVNSASAQKR
Construct: LVNSA(pSer)AQKR (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: 0.46
Immobilized partner concentration (10-6M): 129
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 21

Protein A

Protein: CLASP2
Fragment: 591-600
Site: 14-3-3_pSer596
Modification: (SEP)596
Sequence: VLNTGSDVEE
Construct: VLNTG(pSer)DVEE (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: 0.18
Holdup BI standard deviation: 0.31
Immobilized partner concentration (10-6M): 134
P value: 0.53
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: CLASP2
Fragment: 591-600
Site: 14-3-3_pSer596
Modification: (SEP)596
Sequence: VLNTGSDVEE
Construct: VLNTG(pSer)DVEE (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: -0.05
Holdup BI standard deviation: 0.35
Immobilized partner concentration (10-6M): 129
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))

Experiment series 23

Protein A

Protein: CLASP2
Fragment: 611-620
Site: 14-3-3_pSer616
Modification: (SEP)616
Sequence: RRRYESYGMH
Construct: RRRYE(pSer)YGMH (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: 0.37
Holdup BI standard deviation: 0.19
Immobilized partner concentration (10-6M): 129
P value: 1.66
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.67
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 24

Protein A

Protein: CLASP2
Fragment: 611-620
Site: 14-3-3_pSer616
Modification: (SEP)616
Sequence: RRRYESYGMH
Construct: RRRYE(pSer)YGMH (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: 0.35
Holdup BI standard deviation: 0.13
Immobilized partner concentration (10-6M): 134
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 25

Protein A

Protein: CLASP2
Fragment: 639-648
Site: 14-3-3_pSer644
Modification: (SEP)644
Sequence: SSRNGSIPTY
Construct: SSRNG(pSer)IPTY (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: 0.12
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 134
P value: 3.1
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 26

Protein A

Protein: CLASP2
Fragment: 639-648
Site: 14-3-3_pSer644
Modification: (SEP)644
Sequence: SSRNGSIPTY
Construct: SSRNG(pSer)IPTY (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: 0.08
Holdup BI standard deviation: 0.08
Immobilized partner concentration (10-6M): 129
P value: 1.14
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 27

Protein A

Protein: CLASP2
Fragment: 3-12
Site: 14-3-3_pSer8
Modification: (SEP)8
Sequence: MGDDKSFDDE
Construct: MGDDK(pSer)FDDE (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: -0.17
Holdup BI standard deviation: 0.48
Immobilized partner concentration (10-6M): 134
P value: 0.33
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 28

Protein A

Protein: CLASP2
Fragment: 947-956
Site: 14-3-3_pSer952
Modification: (SEP)952
Sequence: AIQNFSFRSQ
Construct: AIQNF(pSer)FRSQ (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: 0.07
Immobilized partner concentration (10-6M): 134
P value: 0.95
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 29

Protein A

Protein: CLASP2
Fragment: 947-956
Site: 14-3-3_pSer952
Modification: (SEP)952
Sequence: AIQNFSFRSQ
Construct: AIQNF(pSer)FRSQ (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: 0.17
Holdup BI standard deviation: 0.1
Immobilized partner concentration (10-6M): 129
P value: 1.97
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 30

Protein A

Protein: CLASP2
Fragment: 950-959
Site: 14-3-3_pSer955
Modification: (SEP)955
Sequence: NFSFRSQEDM
Construct: NFSFR(pSer)QEDM (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: 0.26
Holdup BI standard deviation: 0.11
Immobilized partner concentration (10-6M): 134
P value: 2.29
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 31

Protein A

Protein: CLASP2
Fragment: 950-959
Site: 14-3-3_pSer955
Modification: (SEP)955
Sequence: NFSFRSQEDM
Construct: NFSFR(pSer)QEDM (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: 0.14
Immobilized partner concentration (10-6M): 129
P value: 0.07
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 32

Protein A

Protein: CLASP2
Fragment: 1007-1016
Site: 14-3-3_pSer1012
Modification: (SEP)1012
Sequence: SMPTHSSPRS
Construct: SMPTH(pSer)SPRS (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: -0.04
Holdup BI standard deviation: 0.84
Immobilized partner concentration (10-6M): 115
P value: 0.04
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 33

Protein A

Protein: CLASP2
Fragment: 1029-1038
Site: 14-3-3_pSer1034
Modification: (SEP)1034
Sequence: SPFNKSALKE
Construct: SPFNK(pSer)ALKE (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: 0.07
Immobilized partner concentration (10-6M): 115
P value: 0.08
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 34

Protein A

Protein: CLASP2
Fragment: 1048-1057
Site: 14-3-3_pSer1053
Modification: (SEP)1053
Sequence: FPDDLSLDHS
Construct: FPDDL(pSer)LDHS (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: -0.03
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 115
P value: 0.51
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 35

Protein A

Protein: CLASP2
Fragment: 347-356
Site: 14-3-3_pSer352
Modification: (SEP)352
Sequence: VAGRVSAGSS
Construct: VAGRV(pSer)AGSS (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: 0.1
Holdup BI standard deviation: 0.17
Immobilized partner concentration (10-6M): 115
P value: 0.48
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 36

Protein A

Protein: CLASP2
Fragment: 355-364
Site: 14-3-3_pSer360
Modification: (SEP)360
Sequence: SSKASSLPGS
Construct: SSKAS(pSer)LPGS (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: 0.77
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 115
P value: 4.07
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.48
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 37

Protein A

Protein: CLASP2
Fragment: 359-368
Site: 14-3-3_pSer364
Modification: (SEP)364
Sequence: SSLPGSLQRS
Construct: SSLPG(pSer)LQRS (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: 0.05
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 115
P value: 1.56
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 38

Protein A

Protein: CLASP2
Fragment: 365-374
Site: 14-3-3_pSer370
Modification: (SEP)370
Sequence: LQRSRSDIDV
Construct: LQRSR(pSer)DIDV (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: 0.06
Holdup BI standard deviation: 0.35
Immobilized partner concentration (10-6M): 115
P value: 0.11
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 39

Protein A

Protein: CLASP2
Fragment: 425-434
Site: 14-3-3_pSer430
Modification: (SEP)430
Sequence: VRAKLSAPLA
Construct: VRAKL(pSer)APLA (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: 0.41
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 115
P value: 5.63
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.8
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 40

Protein A

Protein: CLASP2
Fragment: 450-459
Site: 14-3-3_pSer455
Modification: (SEP)455
Sequence: KMVSQSQPGS
Construct: KMVSQ(pSer)QPGS (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: 0.68
Holdup BI standard deviation: 0.1
Immobilized partner concentration (10-6M): 115
P value: 4.15
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.29
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 41

Protein A

Protein: CLASP2
Fragment: 532-541
Site: 14-3-3_pSer537
Modification: (SEP)537
Sequence: ASRESSRDTS
Construct: ASRES(pSer)RDTS (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: -0.2
Holdup BI standard deviation: 0.79
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))

Experiment series 42

Protein A

Protein: CLASP2
Fragment: 639-648
Site: 14-3-3_pSer644
Modification: (SEP)644
Sequence: SSRNGSIPTY
Construct: SSRNG(pSer)IPTY (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: 0.16
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 115
P value: 4.04
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 43

Protein A

Protein: CLASP2
Fragment: 947-956
Site: 14-3-3_pSer952
Modification: (SEP)952
Sequence: AIQNFSFRSQ
Construct: AIQNF(pSer)FRSQ (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: 0.18
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 115
P value: 2.86
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 44

Protein A

Protein: CLASP2
Fragment: 950-959
Site: 14-3-3_pSer955
Modification: (SEP)955
Sequence: NFSFRSQEDM
Construct: NFSFR(pSer)QEDM (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: -0.26
Holdup BI standard deviation: 0.8
Immobilized partner concentration (10-6M): 115
P value: 0.27
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))