Details of experiments between P63104 and Q7Z3C6
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: YWHAZ
Protein A fragment: 1-245
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3zeta
Protein B protein: ATG9A
Protein B fragment: 736-745
Protein B site: 14-3-3_pSer741
Protein B modification: (SEP)741
Protein B sequence: DESGESAPDE
Protein B construct: DESGE(pSer)APDE (crude synthetic peptide)
Average holdup BI: 0.5
Holdup BI standard deviation: 0.24
Immobilized partner concentration in holdup experiment (10-6M): 132
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 2.25
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: 3.88
Experiment series 2
Protein A protein: YWHAZ
Protein A fragment: 1-245
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3zeta
Protein B protein: ATG9A
Protein B fragment: 750-759
Protein B site: 14-3-3_pSer755
Protein B modification: (SEP)755
Protein B sequence: ARAPQSIPRS
Protein B construct: ARAPQ(pSer)IPRS (crude synthetic peptide)
Average holdup BI: 0.18
Holdup BI standard deviation: 0.09
Immobilized partner concentration in holdup experiment (10-6M): 132
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 2.15
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 3
Protein A protein: YWHAZ
Protein A fragment: 1-245
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3zeta
Protein B protein: ATG9A
Protein B fragment: 9-18
Protein B site: 14-3-3_pSer14
Protein B modification: (SEP)14
Protein B sequence: QRLEASYSDS
Protein B construct: QRLEA(pSer)YSDS (crude synthetic peptide)
Average holdup BI: -0.06
Holdup BI standard deviation: 0.06
Immobilized partner concentration in holdup experiment (10-6M): 132
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 4
Protein A protein: YWHAZ
Protein A fragment: 1-245
Protein A site: 14-3-3
Protein A construct: avi-his6-MBP-TEVsite-14-3-3zeta
Protein B protein: ATG9A
Protein B fragment: 756-765
Protein B site: 14-3-3_pSer761
Protein B modification: (SEP)761
Protein B sequence: IPRSASYPCA
Protein B construct: IPRSA(pSer)YPCA (crude synthetic peptide)
Average holdup BI: 0.2
Holdup BI standard deviation: 0.23
Immobilized partner concentration in holdup experiment (10-6M): 132
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.72
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.