Details of experiments between the fragments Q14160~718-814 and Q96KB5~313-322
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: PBK
Protein B fragment: 313-322
Protein B site: PBM
Protein B sequence: HIVEALETDV
Protein B construct: biotin-ttds-HIVEALETDV
Measured dissociation constant (10-6M): 7.53
Standard deviation of dissociation constant (10-6M): 0.39
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 5.12
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: PBK
Protein B fragment: 313-322
Protein B site: PBM
Protein B sequence: HIVEALETDV
Protein B construct: biotin-ttds-HIVEALETDV
Average holdup BI: 0.46
Immobilized partner concentration in holdup experiment (10-6M): 5.5
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: 5.37
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: PBK
Protein B fragment: 313-322
Protein B site: PBM
Protein B sequence: HIVEALETDV
Protein B construct: biotin-ttds-HIVEALETDV
Average holdup BI: 0.62
Holdup BI standard deviation: 0.03
Normalized holdup BI: 0.65
Normalized holdup BI standard deviation: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 11.6
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: 5.32