Details of experiments between the fragments Q6P0Q8~1102-1193 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: MAST2
Protein A fragment: 1102-1193
Protein A site: PDZ
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
Measured dissociation constant (10-6M): 0.56
Standard deviation of dissociation constant (10-6M): 0.08
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 6.25
Experiment series 2
Protein A protein: MAST2
Protein A fragment: 1102-1193
Protein A site: PDZ
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: 0.56
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: 5.53
Experiment series 3
Protein A protein: MAST2
Protein A fragment: 1102-1193
Protein A site: PDZ
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: 0.85
Holdup BI standard deviation: 0.05
Normalized holdup BI: 0.85
Normalized holdup BI standard deviation: 0.05
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: 2
Measured pKd: 5.76
Experiment series 4
Protein A protein: MAST2
Protein A fragment: 1102-1193
Protein A site: PDZ
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: 0.84
Normalized holdup BI: 0.84
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: 5.55