Details of experiments between the fragments Q14160~718-814 and Q15311~646-655
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: SCRIB
Protein A fragment: 718-814
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: RALBP1
Protein B fragment: 646-655
Protein B site: PBM
Protein B sequence: PSRDRKETSI
Protein B construct: biotin-ttds-PSRDRKETSI
Measured dissociation constant (10-6M): 25.83
Standard deviation of dissociation constant (10-6M): 1.95
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 4.59
Experiment series 2
Protein A protein: SCRIB
Protein A fragment: 718-814
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: RALBP1
Protein B fragment: 646-655
Protein B site: PBM
Protein B sequence: PSRDRKETSI
Protein B construct: biotin-ttds-PSRDRKETSI
Average holdup BI: 0.2
Immobilized partner concentration in holdup experiment (10-6M): 20
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: 4.12
Experiment series 3
Protein A protein: SCRIB
Protein A fragment: 718-814
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: RALBP1
Protein B fragment: 646-655
Protein B site: PBM
Protein B sequence: PSRDRKETSI
Protein B construct: biotin-ttds-PSRDRKETSI
Average holdup BI: 0.43
Holdup BI standard deviation: 0.02
Normalized holdup BI: 0.45
Normalized holdup BI standard deviation: 0.02
Immobilized partner concentration in holdup experiment (10-6M): 28
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: 4
Measured pKd: 4.49
Experiment series 4
Protein A protein: SCRIB
Protein A fragment: 718-814
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: RALBP1
Protein B fragment: 646-655
Protein B site: PBM
Protein B sequence: PSRDRKETSI
Protein B construct: biotin-ttds-PSRDRKETSI
Average holdup BI: 0.5
Normalized holdup BI: 0.5
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: 4.79