Details of experiments between the fragments Q9UBF8~289-298 and P61981~1-247
Note that the results of all experiments are listed, regardless of the modification states of the fragments.



Experiment series 1

Protein A protein: PI4KB
Protein A fragment: 289-298
Protein A site: 14-3-3_pSer294
Protein A modification: (SEP)294
Protein A sequence: LKRTASNPKV
Protein A construct: LKRTA(pSer)NPKV (crude synthetic peptide)

Protein B protein: YWHAG
Protein B fragment: 1-247
Protein B site: 14-3-3
Protein B modification: 100_uM_FCA
Protein B construct: avi-his6-MBP-TEVsite-14-3-3gamma

Measured dissociation constant (10-6M): <10
Immobilized partner concentration in holdup experiment (10-6M): 115
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 1
Measured pKd: 5


Experiment series 2

Protein A protein: PI4KB
Protein A fragment: 289-298
Protein A site: 14-3-3_pSer294
Protein A modification: (SEP)294
Protein A sequence: LKRTASNPKV
Protein A construct: LKRTA(pSer)NPKV (crude synthetic peptide)

Protein B protein: YWHAG
Protein B fragment: 1-247
Protein B site: 14-3-3
Protein B construct: avi-his6-MBP-TEVsite-14-3-3gamma

Measured dissociation constant (10-6M): <10
Immobilized partner concentration in holdup experiment (10-6M): 134
Experiment method: HU Multiplex
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.02.17.
Experimental details: -log10(P Two-sided unpaired T-test (4 vs 4))
Number of measurements: 1
Measured pKd: 5