Details of experiments between the fragments P78352~306-403 and P36809~144-153
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: HPV30-E6
Protein B fragment: 144-153
Protein B site: PBM
Protein B sequence: RHTTSTETAV
Protein B construct: biotin-ttds-RHTTSTETAV
Measured dissociation constant (10-6M): 35.51
Standard deviation of dissociation constant (10-6M): 1.36
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 4.45
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: HPV30-E6
Protein B fragment: 144-153
Protein B site: PBM
Protein B sequence: RHTTSTETAV
Protein B construct: biotin-ttds-RHTTSTETAV
Average holdup BI: 0.16
Immobilized partner concentration in holdup experiment (10-6M): 4
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: 4.75
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: HPV30-E6
Protein B fragment: 144-153
Protein B site: PBM
Protein B sequence: RHTTSTETAV
Protein B construct: biotin-ttds-RHTTSTETAV
Average holdup BI: 0.19
Holdup BI standard deviation: 0
Normalized holdup BI: 0.22
Normalized holdup BI standard deviation: 0
Immobilized partner concentration in holdup experiment (10-6M): 13.4
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: 4.35