Details of experiments between O75122 and P27348
Note that the results of all experiments are listed, regardless of the modification states of the fragments.



Experiment series 1

Protein A protein: CLASP2
Protein A fragment: 355-364
Protein A site: 14-3-3_pSer360
Protein A modification: (SEP)360
Protein A sequence: SSKASSLPGS
Protein A construct: SSKAS(pSer)LPGS (crude synthetic peptide)

Protein B protein: YWHAQ
Protein B fragment: 1-245
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3tau

Average holdup BI: 0.61
Holdup BI standard deviation: 0.02
Immobilized partner concentration in holdup experiment (10-6M): 130
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 9.49
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 5
Measured pKd: 4.08


Experiment series 2

Protein A protein: CLASP2
Protein A fragment: 450-459
Protein A site: 14-3-3_pSer455
Protein A modification: (SEP)455
Protein A sequence: KMVSQSQPGS
Protein A construct: KMVSQ(pSer)QPGS (crude synthetic peptide)

Protein B protein: YWHAQ
Protein B fragment: 1-245
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3tau

Average holdup BI: 0.58
Holdup BI standard deviation: 0.02
Immobilized partner concentration in holdup experiment (10-6M): 130
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 8.81
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 5
Measured pKd: 4.03


Experiment series 3

Protein A protein: CLASP2
Protein A fragment: 365-374
Protein A site: 14-3-3_pSer370
Protein A modification: (SEP)370
Protein A sequence: LQRSRSDIDV
Protein A construct: LQRSR(pSer)DIDV (crude synthetic peptide)

Protein B protein: YWHAQ
Protein B fragment: 1-245
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3tau

Average holdup BI: 0.21
Holdup BI standard deviation: 0.08
Immobilized partner concentration in holdup experiment (10-6M): 130
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 2.93
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 5
Measured pKd: 3.32


Experiment series 4

Protein A protein: CLASP2
Protein A fragment: 359-368
Protein A site: 14-3-3_pSer364
Protein A modification: (SEP)364
Protein A sequence: SSLPGSLQRS
Protein A construct: SSLPG(pSer)LQRS (crude synthetic peptide)

Protein B protein: YWHAQ
Protein B fragment: 1-245
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3tau

Average holdup BI: 0.07
Holdup BI standard deviation: 0.04
Immobilized partner concentration in holdup experiment (10-6M): 130
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 2.23
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 5
The affinity was below the detection threshold of the assay.


Experiment series 5

Protein A protein: CLASP2
Protein A fragment: 425-434
Protein A site: 14-3-3_pSer430
Protein A modification: (SEP)430
Protein A sequence: VRAKLSAPLA
Protein A construct: VRAKL(pSer)APLA (crude synthetic peptide)

Protein B protein: YWHAQ
Protein B fragment: 1-245
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3tau

Average holdup BI: 0.16
Holdup BI standard deviation: 0.08
Immobilized partner concentration in holdup experiment (10-6M): 130
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 2.21
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 5
The affinity was below the detection threshold of the assay.


Experiment series 6

Protein A protein: CLASP2
Protein A fragment: 484-493
Protein A site: 14-3-3_pSer489
Protein A modification: (SEP)489
Protein A sequence: LVNSASAQKR
Protein A construct: LVNSA(pSer)AQKR (crude synthetic peptide)

Protein B protein: YWHAQ
Protein B fragment: 1-245
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3tau

Average holdup BI: 0.26
Holdup BI standard deviation: 0.13
Immobilized partner concentration in holdup experiment (10-6M): 130
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 1.99
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 5
Measured pKd: 3.43


Experiment series 7

Protein A protein: CLASP2
Protein A fragment: 1048-1057
Protein A site: 14-3-3_pSer1053
Protein A modification: (SEP)1053
Protein A sequence: FPDDLSLDHS
Protein A construct: FPDDL(pSer)LDHS (crude synthetic peptide)

Protein B protein: YWHAQ
Protein B fragment: 1-245
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3tau

Average holdup BI: -0.12
Holdup BI standard deviation: 0.1
Immobilized partner concentration in holdup experiment (10-6M): 130
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 1.29
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 5
The affinity was below the detection threshold of the assay.


Experiment series 8

Protein A protein: CLASP2
Protein A fragment: 950-959
Protein A site: 14-3-3_pSer955
Protein A modification: (SEP)955
Protein A sequence: NFSFRSQEDM
Protein A construct: NFSFR(pSer)QEDM (crude synthetic peptide)

Protein B protein: YWHAQ
Protein B fragment: 1-245
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3tau

Average holdup BI: -0.1
Holdup BI standard deviation: 0.11
Immobilized partner concentration in holdup experiment (10-6M): 130
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 1.19
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 5
The affinity was below the detection threshold of the assay.


Experiment series 9

Protein A protein: CLASP2
Protein A fragment: 639-648
Protein A site: 14-3-3_pSer644
Protein A modification: (SEP)644
Protein A sequence: SSRNGSIPTY
Protein A construct: SSRNG(pSer)IPTY (crude synthetic peptide)

Protein B protein: YWHAQ
Protein B fragment: 1-245
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3tau

Average holdup BI: 0.07
Holdup BI standard deviation: 0.07
Immobilized partner concentration in holdup experiment (10-6M): 130
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 1.1
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 5
The affinity was below the detection threshold of the assay.


Experiment series 10

Protein A protein: CLASP2
Protein A fragment: 347-356
Protein A site: 14-3-3_pSer352
Protein A modification: (SEP)352
Protein A sequence: VAGRVSAGSS
Protein A construct: VAGRV(pSer)AGSS (crude synthetic peptide)

Protein B protein: YWHAQ
Protein B fragment: 1-245
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3tau

Average holdup BI: -0.11
Holdup BI standard deviation: 0.14
Immobilized partner concentration in holdup experiment (10-6M): 130
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 1.07
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 5
The affinity was below the detection threshold of the assay.


Experiment series 11

Protein A protein: CLASP2
Protein A fragment: 3-12
Protein A site: 14-3-3_pSer8
Protein A modification: (SEP)8
Protein A sequence: MGDDKSFDDE
Protein A construct: MGDDK(pSer)FDDE (crude synthetic peptide)

Protein B protein: YWHAQ
Protein B fragment: 1-245
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3tau

Average holdup BI: -0.46
Holdup BI standard deviation: 0.96
Immobilized partner concentration in holdup experiment (10-6M): 130
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.78
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 5
The affinity was below the detection threshold of the assay.


Experiment series 12

Protein A protein: CLASP2
Protein A fragment: 591-600
Protein A site: 14-3-3_pSer596
Protein A modification: (SEP)596
Protein A sequence: VLNTGSDVEE
Protein A construct: VLNTG(pSer)DVEE (crude synthetic peptide)

Protein B protein: YWHAQ
Protein B fragment: 1-245
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3tau

Average holdup BI: -0.14
Holdup BI standard deviation: 0.26
Immobilized partner concentration in holdup experiment (10-6M): 130
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.66
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 5
The affinity was below the detection threshold of the assay.


Experiment series 13

Protein A protein: CLASP2
Protein A fragment: 1007-1016
Protein A site: 14-3-3_pSer1012
Protein A modification: (SEP)1012
Protein A sequence: SMPTHSSPRS
Protein A construct: SMPTH(pSer)SPRS (crude synthetic peptide)

Protein B protein: YWHAQ
Protein B fragment: 1-245
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3tau

Average holdup BI: 0.06
Holdup BI standard deviation: 0.13
Immobilized partner concentration in holdup experiment (10-6M): 130
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.43
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 5
The affinity was below the detection threshold of the assay.


Experiment series 14

Protein A protein: CLASP2
Protein A fragment: 1029-1038
Protein A site: 14-3-3_pSer1034
Protein A modification: (SEP)1034
Protein A sequence: SPFNKSALKE
Protein A construct: SPFNK(pSer)ALKE (crude synthetic peptide)

Protein B protein: YWHAQ
Protein B fragment: 1-245
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3tau

Average holdup BI: 0.02
Holdup BI standard deviation: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 130
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.32
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 5
The affinity was below the detection threshold of the assay.


Experiment series 15

Protein A protein: CLASP2
Protein A fragment: 611-620
Protein A site: 14-3-3_pSer616
Protein A modification: (SEP)616
Protein A sequence: RRRYESYGMH
Protein A construct: RRRYE(pSer)YGMH (crude synthetic peptide)

Protein B protein: YWHAQ
Protein B fragment: 1-245
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3tau

Average holdup BI: 0.08
Holdup BI standard deviation: 0.26
Immobilized partner concentration in holdup experiment (10-6M): 130
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.28
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 5
The affinity was below the detection threshold of the assay.


Experiment series 16

Protein A protein: CLASP2
Protein A fragment: 947-956
Protein A site: 14-3-3_pSer952
Protein A modification: (SEP)952
Protein A sequence: AIQNFSFRSQ
Protein A construct: AIQNF(pSer)FRSQ (crude synthetic peptide)

Protein B protein: YWHAQ
Protein B fragment: 1-245
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3tau

Average holdup BI: 0
Holdup BI standard deviation: 0.08
Immobilized partner concentration in holdup experiment (10-6M): 130
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.01
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2021.10.13.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 5))
Number of measurements: 5
The affinity was below the detection threshold of the assay.