Details of experiments between the fragments Q14155~178-251 and P61978~285-299
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: ARHGEF7
Protein A fragment: 178-251
Protein A site: SH3
Protein A construct: avi(biotin)-his6-MBP-TEVsite-SH3
Protein B protein: HNRNPK
Protein B fragment: 285-299
Protein B site: PxxP_4
Protein B sequence: PRRGPPPPPPGRGGR
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.34
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: 4.48
Experiment series 2
Protein A protein: ARHGEF7
Protein A fragment: 178-251
Protein A site: SH3
Protein A construct: his6-MBP-TEVsite-SH3
Protein B protein: HNRNPK
Protein B fragment: 285-299
Protein B site: PxxP_4
Protein B sequence: PRRGPPPPPPGRGGR
Protein B construct: Bio E E Ado Ado-PRRGPPPPPPGRGGR-CONH2
Average holdup BI: 0.1
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: 3.79