Details of experiments between Q12959 and P22459
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: DLG1
Protein A fragment: 314-412
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: KCNA4
Protein B fragment: 644-653
Protein B site: PBM
Protein B sequence: SNAKAVETDV
Protein B construct: biotin-ttds-SNAKAVETDV
Average holdup BI: 0.71
Normalized holdup BI: 0.8
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
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 5.42
Experiment series 2
Protein A protein: DLG1
Protein A fragment: 221-314
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: KCNA4
Protein B fragment: 644-653
Protein B site: PBM
Protein B sequence: SNAKAVETDV
Protein B construct: biotin-ttds-SNAKAVETDV
Average holdup BI: 0.39
Normalized holdup BI: 0.46
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
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 4.72
Experiment series 3
Protein A protein: DLG1
Protein A fragment: 457-570
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: KCNA4
Protein B fragment: 644-653
Protein B site: PBM
Protein B sequence: SNAKAVETDV
Protein B construct: biotin-ttds-SNAKAVETDV
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
PUBMED ID of publication: 36115835
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 4
Protein A protein: DLG1
Protein A fragment: 221-314
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: KCNA4
Protein B fragment: 644-653
Protein B site: PBM
Protein B sequence: SNAKAVETDV
Protein B construct: Biotin-ado-ado-SNAKAVETDV
Average holdup BI: 0.42
Normalized holdup BI: 0.49
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: 4.78
Experiment series 5
Protein A protein: DLG1
Protein A fragment: 314-412
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: KCNA4
Protein B fragment: 644-653
Protein B site: PBM
Protein B sequence: SNAKAVETDV
Protein B construct: Biotin-ado-ado-SNAKAVETDV
Average holdup BI: 0.75
Normalized holdup BI: 0.83
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: 5.52
Experiment series 6
Protein A protein: DLG1
Protein A fragment: 457-570
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: KCNA4
Protein B fragment: 644-653
Protein B site: PBM
Protein B sequence: SNAKAVETDV
Protein B construct: Biotin-ado-ado-SNAKAVETDV
Average holdup BI: -0.04
Normalized holdup BI: -0.06
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
The affinity was below the detection threshold of the assay.