Details of experiments between the fragments P49418~615-695 and P30260~396-410
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: AMPH
Protein A fragment: 615-695
Protein A site: SH3
Protein A construct: avi(biotin)-his6-MBP-TEVsite-SH3
Protein B protein: CDC27
Protein B fragment: 396-410
Protein B site: PxxP_1
Protein B sequence: LKMKFPPKIPNRKTK
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.61
Holdup BI standard deviation: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.06.29.
Number of measurements: 4
Measured pKd: 5
Experiment series 2
Protein A protein: AMPH
Protein A fragment: 615-695
Protein A site: SH3
Protein A construct: his6-MBP-TEVsite-SH3
Protein B protein: CDC27
Protein B fragment: 396-410
Protein B site: PxxP_1
Protein B sequence: LKMKFPPKIPNRKTK
Protein B construct: Bio E E Ado Ado-LKMKFPPKIPNRKTK-CONH2
Average holdup BI: 0.47
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/16
Number of measurements: 1
Measured pKd: 4.74