Details of experiments between the fragments P49418~615-695 and Q9BRD0~219-233
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: BUD13
Protein B fragment: 219-233
Protein B site: PxxP_2
Protein B sequence: RHDSPDPSPPRRARH
Protein B construct: Biotinylated 15-mer peptide (crude)

Average holdup BI: 0.26
Holdup BI standard deviation: 0.01
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: 4.31


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: BUD13
Protein B fragment: 219-233
Protein B site: PxxP_2
Protein B sequence: RHDSPDPSPPRRARH
Protein B construct: Bio E E Ado Ado-RHDSPDPSPPRRARH-CONH2

Average holdup BI: 0.15
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: 3.99