Details of experiments between Q9UPQ7 and Q9UDY2
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: PDZRN3
Protein A fragment: 1057-1066
Protein A site: PBM
Protein A sequence: YNSFLSVTTV
Protein A construct: Biotin-Ahx-YNSFLSVTTV
Protein B protein: TJP2
Protein B fragment: 24-128
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
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 2
Protein A protein: PDZRN3
Protein A fragment: 245-339
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: TJP2
Protein B fragment: 1181-1190
Protein B site: PBM
Protein B sequence: QSARYRDTEL
Protein B construct: biotin-ttds-QSARYRDTEL
Average holdup BI: 0.24
Immobilized partner concentration in holdup experiment (10-6M): 10
Experiment method: SAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normal layout
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 4.54
Experiment series 3
Protein A protein: PDZRN3
Protein A fragment: 405-515
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: TJP2
Protein B fragment: 1181-1190
Protein B site: PBM
Protein B sequence: QSARYRDTEL
Protein B construct: biotin-ttds-QSARYRDTEL
Average holdup BI: -0.13
Immobilized partner concentration in holdup experiment (10-6M): 10
Experiment method: SAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normal 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: PDZRN3
Protein A fragment: 245-339
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: TJP2
Protein B fragment: 1181-1190
Protein B site: PBM
Protein B sequence: QSARYRDTEL
Protein B construct: biotin-ttds-QSARYRDTEL
Average holdup BI: 0.41
Normalized holdup BI: 0.43
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: 4.67