Details of experiments between the fragments P78352~306-403 and P26554~142-151
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: DLG4
Protein A fragment: 306-403
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: HPV51-E6
Protein B fragment: 142-151
Protein B site: PBM
Protein B sequence: RTRQRNETQV
Protein B construct: biotin-ttds-RTRQRNETQV
Measured dissociation constant (10-6M): 9.54
Standard deviation of dissociation constant (10-6M): 0.91
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 5.02
Experiment series 2
Protein A protein: DLG4
Protein A fragment: 306-403
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: HPV51-E6
Protein B fragment: 142-151
Protein B site: PBM
Protein B sequence: RTRQRNETQV
Protein B construct: biotin-ttds-RTRQRNETQV
Average holdup BI: 0.25
Immobilized partner concentration in holdup experiment (10-6M): 7.7
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: 4.69
Experiment series 3
Protein A protein: DLG4
Protein A fragment: 306-403
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: HPV51-E6
Protein B fragment: 142-151
Protein B site: PBM
Protein B sequence: RTRQRNETQV
Protein B construct: biotin-ttds-RTRQRNETQV
Average holdup BI: 0.45
Holdup BI standard deviation: 0.02
Normalized holdup BI: 0.52
Normalized holdup BI standard deviation: 0.02
Immobilized partner concentration in holdup experiment (10-6M): 21.5
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: 4.75