Details of experiments between the fragments Q96L92~40-141 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: SNX27
Protein A fragment: 40-141
Protein A site: PDZ
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): 71.14
Standard deviation of dissociation constant (10-6M): 11.23
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 4.15
Experiment series 2
Protein A protein: SNX27
Protein A fragment: 40-141
Protein A site: PDZ
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: 0.4
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: 4.55
Experiment series 3
Protein A protein: SNX27
Protein A fragment: 40-141
Protein A site: PDZ
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: 0.31
Holdup BI standard deviation: 0.03
Normalized holdup BI: 0.31
Normalized holdup BI standard deviation: 0.03
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: 3
Measured pKd: 4.23
Experiment series 4
Protein A protein: SNX27
Protein A fragment: 40-141
Protein A site: PDZ
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: 0.34
Normalized holdup BI: 0.34
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: 4.49