Details of experiments between the fragments P50570~826-840 and Q14155~178-251
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: DNM2
Protein A fragment: 826-840
Protein A site: PxxP_4
Protein A sequence: PAPPQIPSRPVRIPP
Protein A construct: Biotinylated 15-mer peptide (crude)
Protein B protein: ARHGEF7
Protein B fragment: 178-251
Protein B site: SH3
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: 0.41
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: 2
Measured pKd: 4.64
Experiment series 2
Protein A protein: DNM2
Protein A fragment: 826-840
Protein A site: PxxP_4
Protein A sequence: PAPPQIPSRPVRIPP
Protein A construct: Bio E E Ado Ado-PAPPQIPSRPVRIPP-CONH2
Protein B protein: ARHGEF7
Protein B fragment: 178-251
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.38
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/17
Number of measurements: 1
Measured pKd: 4.57