Details of experiments between the fragments P49418~615-695 and Q92619~1106-1120
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: ARHGAP45
Protein B fragment: 1106-1120
Protein B site: PxxP_1
Protein B sequence: NNVLQAPLPPMRLRG
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.3
Holdup BI standard deviation: 0.02
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.06.29.
Number of measurements: 2
Measured pKd: 4.41
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: ARHGAP45
Protein B fragment: 1106-1120
Protein B site: PxxP_1
Protein B sequence: NNVLQAPLPPMRLRG
Protein B construct: Bio E E Ado Ado-NNVLQAPLPPMRLRG-CONH2
Average holdup BI: 0.23
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: 4.23