Interaction Details


Details of experiments between Q7Z460 and P63104
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: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta
Average holdup BI: 0.03
Holdup BI standard deviation: 0.16
Immobilized partner concentration (10-6M): 132
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 2

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: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta
Average holdup BI: 0.02
Holdup BI standard deviation: 0.73
Immobilized partner concentration (10-6M): 132
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 3

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: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta
Average holdup BI: 0.16
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 132
P value: 4.41
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: CLASP1
Fragment: 593-602
Site: 14-3-3_pSer598
Modification: (SEP)598
Sequence: GSLQRSRSDI
Construct: GSLQR(pSer)RSDI (crude synthetic peptide)

Protein B

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta
Average holdup BI: 0.07
Holdup BI standard deviation: 0.09
Immobilized partner concentration (10-6M): 132
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 5

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: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta
Average holdup BI: 0.02
Holdup BI standard deviation: 0.09
Immobilized partner concentration (10-6M): 132
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 6

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: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta
Average holdup BI: 0.32
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 132
P value: 4.37
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.55
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: 682-691
Site: 14-3-3_pSer687
Modification: (SEP)687
Sequence: KVVSQSQPGS
Construct: KVVSQ(pSer)QPGS (crude synthetic peptide)

Protein B

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta
Average holdup BI: 0.62
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 132
P value: 6.73
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.09
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: 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: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta
Measured dissociation constant (10-6M): 101
Standard deviation of dissociation constant (10-6M): 15
Experiment method: competitive Fluorescence Polarization
Details of affinity fitting: Competitive binding equation in ProFit
Date of measurement: 2022-01-19
Number of measurements: 3
Measured pKd: 3.99
DOI: 10.1101/2022.12.08.519103

Experiment series 9

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: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta
Measured dissociation constant (10-6M): 109
Standard deviation of dissociation constant (10-6M): 18
Experiment method: competitive Fluorescence Polarization
Details of affinity fitting: Competitive binding equation in ProFit
Date of measurement: 2022-02-10
Number of measurements: 3
Measured pKd: 3.96
DOI: 10.1101/2022.12.08.519103

Experiment series 10

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: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta
Average holdup BI: 0.02
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 132
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 11

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: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta
Average holdup BI: 0
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 132
P value: 0.17
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 12

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: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta
Average holdup BI: 0.66
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 132
P value: 10.5
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.17
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: CLASP1
Fragment: 277-286
Site: 14-3-3_pThr282
Modification: (TPO)282
Sequence: RLGSSTLGSK
Construct: RLGSS(pThr)LGSK (crude synthetic peptide)

Protein B

Protein: YWHAZ
Fragment: 1-245
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3zeta
Average holdup BI: 0.45
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 132
P value: 6.87
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.79
DOI: 10.1101/2022.12.08.519103
Experimental Details

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