Interaction Details


Details of experiments between Q7Z460 and Q04917
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: 563-572
Site: 14-3-3_pSer568
Modification: (SEP)568
Sequence: LNRPLSAKRS
Construct: LNRPL(pSer)AKRS (crude synthetic peptide)

Protein B

Protein: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta
Average holdup BI: -0.47
Holdup BI standard deviation: 0.93
Immobilized partner concentration (10-6M): 130
P value: 0.71
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: 589-598
Site: 14-3-3_pSer594
Modification: (SEP)594
Sequence: VSTTGSLQRS
Construct: VSTTG(pSer)LQRS (crude synthetic peptide)

Protein B

Protein: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta
Average holdup BI: 0.18
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 130
P value: 4.59
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: 593-602
Site: 14-3-3_pSer598
Modification: (SEP)598
Sequence: GSLQRSRSDI
Construct: GSLQR(pSer)RSDI (crude synthetic peptide)

Protein B

Protein: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta
Average holdup BI: 0.01
Holdup BI standard deviation: 0.08
Immobilized partner concentration (10-6M): 130
P value: 0.1
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: 628-637
Site: 14-3-3_pSer633
Modification: (SEP)633
Sequence: ALPPGSYASL
Construct: ALPPG(pSer)YASL (crude synthetic peptide)

Protein B

Protein: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta
Average holdup BI: 0.04
Holdup BI standard deviation: 0.1
Immobilized partner concentration (10-6M): 130
P value: 0.34
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: 631-640
Site: 14-3-3_pSer636
Modification: (SEP)636
Sequence: PGSYASLGRI
Construct: PGSYA(pSer)LGRI (crude synthetic peptide)

Protein B

Protein: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta
Average holdup BI: 0.44
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 130
P value: 5.75
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 3.77
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: 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: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta
Measured dissociation constant (10-6M): 42
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: 2022-01-19
Number of measurements: 3
Measured pKd: 4.38
DOI: 10.1101/2022.12.08.519103

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: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta
Average holdup BI: 0.71
Holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 130
P value: 6.05
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.28
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: 790-799
Site: 14-3-3_pSer795
Modification: (SEP)795
Sequence: AMRVLSTSTD
Construct: AMRVL(pSer)TSTD (crude synthetic peptide)

Protein B

Protein: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta
Average holdup BI: 0.04
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 130
P value: 0.65
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: 792-801
Site: 14-3-3_pSer797
Modification: (SEP)797
Sequence: RVLSTSTDLE
Construct: RVLST(pSer)TDLE (crude synthetic peptide)

Protein B

Protein: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta
Average holdup BI: 0.01
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 130
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 10

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: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta
Average holdup BI: 0.69
Holdup BI standard deviation: 0.08
Immobilized partner concentration (10-6M): 130
P value: 6.23
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.24
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: 277-286
Site: 14-3-3_pThr282
Modification: (TPO)282
Sequence: RLGSSTLGSK
Construct: RLGSS(pThr)LGSK (crude synthetic peptide)

Protein B

Protein: YWHAH
Fragment: 1-246
Site: 14-3-3
Construct: avi-his6-MBP-TEVsite-14-3-3eta
Average holdup BI: 0.63
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 130
P value: 6.79
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021-10-13
Number of measurements: 5
Measured pKd: 4.11
DOI: 10.1101/2022.12.08.519103
Experimental Details

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