Interaction Details


Details of experiments between Q7Z460 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: CLASP1
Fragment: 1191-1200
Site: 14-3-3_pSer1196
Modification: (SEP)1196
Sequence: KFSFRSQEDL
Construct: KFSFR(pSer)QEDL (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: -0.29
Holdup BI standard deviation: 0.21
Immobilized partner concentration (10-6M): 134
P value: 1.77
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: CLASP1
Fragment: 1191-1200
Site: 14-3-3_pSer1196
Modification: (SEP)1196
Sequence: KFSFRSQEDL
Construct: KFSFR(pSer)QEDL (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: 0.15
Holdup BI standard deviation: 0.18
Immobilized partner concentration (10-6M): 129
P value: 0.83
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: CLASP1
Fragment: 563-572
Site: 14-3-3_pSer568
Modification: (SEP)568
Sequence: LNRPLSAKRS
Construct: LNRPL(pSer)AKRS (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: 0.65
Immobilized partner concentration (10-6M): 134
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 4

Protein A

Protein: CLASP1
Fragment: 563-572
Site: 14-3-3_pSer568
Modification: (SEP)568
Sequence: LNRPLSAKRS
Construct: LNRPL(pSer)AKRS (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: 0.15
Holdup BI standard deviation: 0.59
Immobilized partner concentration (10-6M): 129
P value: 0.2
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 5

Protein A

Protein: CLASP1
Fragment: 589-598
Site: 14-3-3_pSer594
Modification: (SEP)594
Sequence: VSTTGSLQRS
Construct: VSTTG(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: 0.18
Holdup BI standard deviation: 0.09
Immobilized partner concentration (10-6M): 134
P value: 2.1
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 6

Protein A

Protein: CLASP1
Fragment: 589-598
Site: 14-3-3_pSer594
Modification: (SEP)594
Sequence: VSTTGSLQRS
Construct: VSTTG(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: 0.22
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 129
P value: 5.16
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: CLASP1
Fragment: 593-602
Site: 14-3-3_pSer598
Modification: (SEP)598
Sequence: GSLQRSRSDI
Construct: GSLQR(pSer)RSDI (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: 0.04
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 134
P value: 0.58
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: CLASP1
Fragment: 593-602
Site: 14-3-3_pSer598
Modification: (SEP)598
Sequence: GSLQRSRSDI
Construct: GSLQR(pSer)RSDI (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: 0.08
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 9

Protein A

Protein: CLASP1
Fragment: 628-637
Site: 14-3-3_pSer633
Modification: (SEP)633
Sequence: ALPPGSYASL
Construct: ALPPG(pSer)YASL (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: 0.34
Holdup BI standard deviation: 0.3
Immobilized partner concentration (10-6M): 134
P value: 1.04
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 10

Protein A

Protein: CLASP1
Fragment: 628-637
Site: 14-3-3_pSer633
Modification: (SEP)633
Sequence: ALPPGSYASL
Construct: ALPPG(pSer)YASL (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: 0.09
Immobilized partner concentration (10-6M): 129
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 11

Protein A

Protein: CLASP1
Fragment: 631-640
Site: 14-3-3_pSer636
Modification: (SEP)636
Sequence: PGSYASLGRI
Construct: PGSYA(pSer)LGRI (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: 0.51
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 129
P value: 5.96
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.91
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 12

Protein A

Protein: CLASP1
Fragment: 631-640
Site: 14-3-3_pSer636
Modification: (SEP)636
Sequence: PGSYASLGRI
Construct: PGSYA(pSer)LGRI (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: 0.49
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 134
P value: 5.95
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 3.85
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: CLASP1
Fragment: 682-691
Site: 14-3-3_pSer687
Modification: (SEP)687
Sequence: KVVSQSQPGS
Construct: KVVSQ(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: 0.06
Immobilized partner concentration (10-6M): 129
P value: 6.62
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.53
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: CLASP1
Fragment: 682-691
Site: 14-3-3_pSer687
Modification: (SEP)687
Sequence: KVVSQSQPGS
Construct: biotin-ttds-KVVSQ(pSer)QPGS (HPLC-purified synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Measured dissociation constant (10-6M): 25
Standard deviation of dissociation constant (10-6M): 4
Experiment method: competitive Fluorescence Polarization
Details of affinity fitting: Competitive binding equation in ProFit
Date of measurement: 2022-01-10
Number of measurements: 3
Measured pKd: 4.42
DOI: 10.1101/2022.12.08.519103

Experiment series 15

Protein A

Protein: CLASP1
Fragment: 682-691
Site: 14-3-3_pSer687
Modification: (SEP)687
Sequence: KVVSQSQPGS
Construct: KVVSQ(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: 0.76
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 134
P value: 5.58
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.38
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: CLASP1
Fragment: 682-691
Site: 14-3-3_pSer687
Modification: (SEP)687
Sequence: KVVSQSQPGS
Construct: biotin-ttds-KVVSQ(pSer)QPGS (HPLC-purified synthetic peptide)

Protein B

Protein: YWHAG
Fragment: 1-247
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3gamma
Measured dissociation constant (10-6M): 48
Standard deviation of dissociation constant (10-6M): 4.5
Experiment method: competitive Fluorescence Polarization
Details of affinity fitting: Competitive binding equation in ProFit
Date of measurement: 2022-01-18
Number of measurements: 3
Measured pKd: 4.32
DOI: 10.1101/2022.12.08.519103

Experiment series 17

Protein A

Protein: CLASP1
Fragment: 790-799
Site: 14-3-3_pSer795
Modification: (SEP)795
Sequence: AMRVLSTSTD
Construct: AMRVL(pSer)TSTD (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: 0.14
Immobilized partner concentration (10-6M): 134
P value: 1.31
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 18

Protein A

Protein: CLASP1
Fragment: 790-799
Site: 14-3-3_pSer795
Modification: (SEP)795
Sequence: AMRVLSTSTD
Construct: AMRVL(pSer)TSTD (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: 0.05
Immobilized partner concentration (10-6M): 129
P value: 0.15
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 19

Protein A

Protein: CLASP1
Fragment: 792-801
Site: 14-3-3_pSer797
Modification: (SEP)797
Sequence: RVLSTSTDLE
Construct: RVLST(pSer)TDLE (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: 0.04
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 134
P value: 1.96
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: CLASP1
Fragment: 792-801
Site: 14-3-3_pSer797
Modification: (SEP)797
Sequence: RVLSTSTDLE
Construct: RVLST(pSer)TDLE (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: 0.04
Immobilized partner concentration (10-6M): 129
P value: 0.61
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: CLASP1
Fragment: 1079-1088
Site: 14-3-3_pThr1084
Modification: (TPO)1084
Sequence: RTSPLTSPTN
Construct: RTSPL(pThr)SPTN (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: 0.77
Holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 129
P value: 12.53
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.41
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 22

Protein A

Protein: CLASP1
Fragment: 1079-1088
Site: 14-3-3_pThr1084
Modification: (TPO)1084
Sequence: RTSPLTSPTN
Construct: RTSPL(pThr)SPTN (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: 0.75
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 134
P value: 10.95
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.35
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: CLASP1
Fragment: 277-286
Site: 14-3-3_pThr282
Modification: (TPO)282
Sequence: RLGSSTLGSK
Construct: RLGSS(pThr)LGSK (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: 0.74
Holdup BI standard deviation: 0.08
Immobilized partner concentration (10-6M): 129
P value: 6.38
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.33
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: CLASP1
Fragment: 277-286
Site: 14-3-3_pThr282
Modification: (TPO)282
Sequence: RLGSSTLGSK
Construct: RLGSS(pThr)LGSK (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: 0.66
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 134
P value: 6.51
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.16
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: CLASP1
Fragment: 1191-1200
Site: 14-3-3_pSer1196
Modification: (SEP)1196
Sequence: KFSFRSQEDL
Construct: KFSFR(pSer)QEDL (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: 0.12
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 115
P value: 1.87
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 26

Protein A

Protein: CLASP1
Fragment: 563-572
Site: 14-3-3_pSer568
Modification: (SEP)568
Sequence: LNRPLSAKRS
Construct: LNRPL(pSer)AKRS (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: 0.26
Immobilized partner concentration (10-6M): 115
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 27

Protein A

Protein: CLASP1
Fragment: 589-598
Site: 14-3-3_pSer594
Modification: (SEP)594
Sequence: VSTTGSLQRS
Construct: VSTTG(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: 0.12
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 115
P value: 2.32
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: CLASP1
Fragment: 593-602
Site: 14-3-3_pSer598
Modification: (SEP)598
Sequence: GSLQRSRSDI
Construct: GSLQR(pSer)RSDI (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: 0.23
Holdup BI standard deviation: 0.17
Immobilized partner concentration (10-6M): 115
P value: 1.32
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: CLASP1
Fragment: 628-637
Site: 14-3-3_pSer633
Modification: (SEP)633
Sequence: ALPPGSYASL
Construct: ALPPG(pSer)YASL (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: 1.02
Immobilized partner concentration (10-6M): 115
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 30

Protein A

Protein: CLASP1
Fragment: 631-640
Site: 14-3-3_pSer636
Modification: (SEP)636
Sequence: PGSYASLGRI
Construct: PGSYA(pSer)LGRI (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: 0.53
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 115
P value: 4.51
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4
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: CLASP1
Fragment: 682-691
Site: 14-3-3_pSer687
Modification: (SEP)687
Sequence: KVVSQSQPGS
Construct: KVVSQ(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: 0.69
Holdup BI standard deviation: 0.09
Immobilized partner concentration (10-6M): 115
P value: 2.5
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.3
DOI: 10.1101/2022.12.08.519103
Experimental Details

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

Experiment series 32

Protein A

Protein: CLASP1
Fragment: 718-727
Site: 14-3-3_pSer723
Modification: (SEP)723
Sequence: SKIPRSQGCS
Construct: SKIPR(pSer)QGCS (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: 0.38
Immobilized partner concentration (10-6M): 115
P value: 0.28
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: CLASP1
Fragment: 790-799
Site: 14-3-3_pSer795
Modification: (SEP)795
Sequence: AMRVLSTSTD
Construct: AMRVL(pSer)TSTD (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: 0.12
Holdup BI standard deviation: 0.33
Immobilized partner concentration (10-6M): 115
P value: 0.31
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: CLASP1
Fragment: 792-801
Site: 14-3-3_pSer797
Modification: (SEP)797
Sequence: RVLSTSTDLE
Construct: RVLST(pSer)TDLE (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: 0.07
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 115
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 35

Protein A

Protein: CLASP1
Fragment: 1079-1088
Site: 14-3-3_pThr1084
Modification: (TPO)1084
Sequence: RTSPLTSPTN
Construct: RTSPL(pThr)SPTN (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: 0.7
Holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 115
P value: 12.42
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-02-17
Number of measurements: 4
Measured pKd: 4.33
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: CLASP1
Fragment: 277-286
Site: 14-3-3_pThr282
Modification: (TPO)282
Sequence: RLGSSTLGSK
Construct: RLGSS(pThr)LGSK (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: 0.61
Holdup BI standard deviation: 0.16
Immobilized partner concentration (10-6M): 115
P value: 1.86
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))