Details of experiments between Q7Z6J2 and Q9UPQ7
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: GRASP
Protein A fragment: 386-395
Protein A site: PBM
Protein A sequence: RSLEEEESQL
Protein A construct: biotin-ttds-RSLEEEESQL
Protein B protein: PDZRN3
Protein B fragment: 245-339
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.11
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: 4.11
Experiment series 2
Protein A protein: GRASP
Protein A fragment: 386-395
Protein A site: PBM
Protein A sequence: RSLEEEESQL
Protein A construct: biotin-ttds-RSLEEEESQL
Protein B protein: PDZRN3
Protein B fragment: 405-515
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0
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 3
Protein A protein: GRASP
Protein A fragment: 386-395
Protein A site: PBM
Protein A sequence: RSLEEEESQL
Protein A construct: biotin-ttds-RSLEEEESQL
Protein B protein: PDZRN3
Protein B fragment: 245-339
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.22
Normalized holdup BI: 0.23
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: 4.24
Experiment series 4
Protein A protein: GRASP
Protein A fragment: 96-193
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: PDZRN3
Protein B fragment: 1057-1066
Protein B site: PBM
Protein B sequence: YNSFLSVTTV
Protein B construct: Biotin-Ahx-YNSFLSVTTV
Average holdup BI: 0.2
Normalized holdup BI: 0.2
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: 4.15