Details of experiments between P27348 and Q9UJF2
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: YWHAQ
Protein A fragment: 1-245
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3tau
Protein B protein: RASAL2
Protein B fragment: 612-621
Protein B site: 14-3-3_pThr617
Protein B modification: (TPO)617
Protein B sequence: ITKSLTNPTP
Protein B construct: ITKSL(pThr)NPTP (crude synthetic peptide)
Average holdup BI: 0.67
Holdup BI standard deviation: 0.03
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): 7.26
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: 4.2
Experiment series 2
Protein A protein: YWHAQ
Protein A fragment: 1-245
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3tau
Protein B protein: RASAL2
Protein B fragment: 129-138
Protein B site: 14-3-3_pSer134
Protein B modification: (SEP)134
Protein B sequence: LRRTVSVPSE
Protein B construct: LRRTV(pSer)VPSE (crude synthetic peptide)
Average holdup BI: 0.79
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): 7.21
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
Measured pKd: 4.45
Experiment series 3
Protein A protein: YWHAQ
Protein A fragment: 1-245
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3tau
Protein B protein: RASAL2
Protein B fragment: 844-853
Protein B site: 14-3-3_pThr849
Protein B modification: (TPO)849
Protein B sequence: FSRRHTVPDR
Protein B construct: FSRRH(pThr)VPDR (crude synthetic peptide)
Average holdup BI: 0.84
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): 3.83
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: 4.62
Experiment series 4
Protein A protein: YWHAQ
Protein A fragment: 1-245
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3tau
Protein B protein: RASAL2
Protein B fragment: 836-845
Protein B site: 14-3-3_pSer841
Protein B modification: (SEP)841
Protein B sequence: KRSTQSEDFS
Protein B construct: KRSTQ(pSer)EDFS (crude synthetic peptide)
Average holdup BI: 0.39
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): 2.61
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: 3.7
Experiment series 5
Protein A protein: YWHAQ
Protein A fragment: 1-245
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3tau
Protein B protein: RASAL2
Protein B fragment: 84-93
Protein B site: 14-3-3_pSer89
Protein B modification: (SEP)89
Protein B sequence: LKESRSHESL
Protein B construct: LKESR(pSer)HESL (crude synthetic peptide)
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): 0.42
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: YWHAQ
Protein A fragment: 1-245
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3tau
Protein B protein: RASAL2
Protein B fragment: 936-945
Protein B site: 14-3-3_pThr941
Protein B modification: (TPO)941
Protein B sequence: IQRQQTQQVQ
Protein B construct: IQRQQ(pThr)QQVQ (crude synthetic peptide)
Average holdup BI: 0.05
Holdup BI standard deviation: 0.16
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): 0.27
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 7
Protein A protein: YWHAQ
Protein A fragment: 1-245
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3tau
Protein B protein: RASAL2
Protein B fragment: 734-743
Protein B site: 14-3-3_pSer739
Protein B modification: (SEP)739
Protein B sequence: TGSQLSITQV
Protein B construct: TGSQL(pSer)ITQV (crude synthetic peptide)
Average holdup BI: 0.02
Holdup BI standard deviation: 0.06
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): 0.23
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 8
Protein A protein: YWHAQ
Protein A fragment: 1-245
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3tau
Protein B protein: RASAL2
Protein B fragment: 882-891
Protein B site: 14-3-3_pSer887
Protein B modification: (SEP)887
Protein B sequence: AKAPPSLPHS
Protein B construct: AKAPP(pSer)LPHS (crude synthetic peptide)
Average holdup BI: 0.03
Holdup BI standard deviation: 0.2
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): 0.12
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.