Details of experiments between the fragments Q5T2W1~2-107 and Q13574~919-928
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: NHERF3
Protein A fragment: 2-107
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: DGKZ
Protein B fragment: 919-928
Protein B site: PBM
Protein B sequence: IQREDQETAV
Protein B construct: biotin-ttds-IQREDQETAV
Average holdup BI: 0.73
Immobilized partner concentration in holdup experiment (10-6M): 6
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: 5.95
Experiment series 2
Protein A protein: NHERF3
Protein A fragment: 2-107
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: DGKZ
Protein B fragment: 919-928
Protein B site: PBM
Protein B sequence: IQREDQETAV
Protein B construct: biotin-ttds-IQREDQETAV
Average holdup BI: 0.57
Holdup BI standard deviation: 0.03
Normalized holdup BI: 0.6
Normalized holdup BI standard deviation: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 13.5
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: 3
Measured pKd: 5.14
Experiment series 3
Protein A protein: NHERF3
Protein A fragment: 2-107
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: DGKZ
Protein B fragment: 919-928
Protein B site: PBM
Protein B sequence: IQREDQETAV
Protein B construct: biotin-ttds-IQREDQETAV
Average holdup BI: 0.63
Normalized holdup BI: 0.67
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: 5.11