Details of experiments between the fragments O15085~41-126 and Q7Z628~587-596
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: ARHGEF11
Protein A fragment: 41-126
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: NET1
Protein B fragment: 587-596
Protein B site: PBM
Protein B sequence: SGGKRKETLV
Protein B construct: biotin-ttds-SGGKRKETLV
Average holdup BI: 0.18
Holdup BI standard deviation: 0.04
Normalized holdup BI: 0.22
Normalized holdup BI standard deviation: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 17.9
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
PUBMED ID of publication: 36115835
Number of measurements: 2
Measured pKd: 4.23
Experiment series 2
Protein A protein: ARHGEF11
Protein A fragment: 41-126
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: NET1
Protein B fragment: 587-596
Protein B site: PBM
Protein B sequence: SGGKRKETLV
Protein B construct: biotin-ttds-SGGKRKETLV
Average holdup BI: -0
Normalized holdup BI: -0
Immobilized partner concentration in holdup experiment (10-6M): 17.9
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
Experimental details: not reported internal standards of holdup layout #5
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 3
Protein A protein: ARHGEF11
Protein A fragment: 41-126
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: NET1
Protein B fragment: 587-596
Protein B site: PBM
Protein B sequence: SGGKRKETLV
Protein B construct: Biotin-ado-ado-SGGKRKETLV
Average holdup BI: 0.05
Normalized holdup BI: 0.08
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
Experimental details: Crude peptide synthesis product. The peptide concentration in the affinity conversion step may differ from reality. Not reported internal standards of holdup layout #6
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 3.68