Details of experiments between A4D2P6 and P25054
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: GRID2IP
Protein A fragment: 272-370
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: APC
Protein B fragment: 2834-2843
Protein B site: PBM
Protein B sequence: HSGSYLVTSV
Protein B construct: Biotin-Ahx-HSGSYLVTSV
Average holdup BI: 0.27
Normalized holdup BI: 0.3
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: The peptide stock solution had solubility issues. The peptide concentration in the affinity conversion step may differ from reality causing a systematic error in reported affinities.
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 4.41
Experiment series 2
Protein A protein: GRID2IP
Protein A fragment: 1-87
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: APC
Protein B fragment: 2834-2843
Protein B site: PBM
Protein B sequence: HSGSYLVTSV
Protein B construct: Biotin-Ahx-HSGSYLVTSV
Average holdup BI: -0.02
Normalized holdup BI: -0.02
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: The peptide stock solution had solubility issues. The peptide concentration in the affinity conversion step may differ from reality causing a systematic error in reported affinities.
PUBMED ID of publication: 36115835
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 3
Protein A protein: GRID2IP
Protein A fragment: 272-370
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: APC
Protein B fragment: 2834-2843
Protein B site: PBM
Protein B sequence: HSGSYLVTSV
Protein B construct: Biotin-Ahx-HSGSYLVTSV
Average holdup BI: 0.38
Normalized holdup BI: 0.38
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: not reported internal standards of holdup layout #6
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 4.57
Experiment series 4
Protein A protein: GRID2IP
Protein A fragment: 1-87
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: APC
Protein B fragment: 2834-2843
Protein B site: PBM
Protein B sequence: HSGSYLVTSV
Protein B construct: Biotin-Ahx-HSGSYLVTSV
Average holdup BI: 0.02
Normalized holdup BI: 0.02
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: not reported internal standards of holdup layout #6
PUBMED ID of publication: 36115835
Number of measurements: 1
The affinity was below the detection threshold of the assay.