Details of experiments between the fragments P06427~140-149 and P78352~306-403
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: HPV33-E6
Protein A fragment: 140-149
Protein A site: PBM
Protein A sequence: SRSRRRETAL
Protein A construct: biotin-ttds-SRSRRRETAL
Protein B protein: DLG4
Protein B fragment: 306-403
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ
Measured dissociation constant (10-6M): 3.91
Standard deviation of dissociation constant (10-6M): 0.34
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 5.41
Experiment series 2
Protein A protein: HPV33-E6
Protein A fragment: 140-149
Protein A site: PBM
Protein A sequence: SRSRRRETAL
Protein A construct: biotin-ttds-SRSRRRETAL
Protein B protein: DLG4
Protein B fragment: 306-403
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.47
Immobilized partner concentration in holdup experiment (10-6M): 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: 5.24
Experiment series 3
Protein A protein: HPV33-E6
Protein A fragment: 140-149
Protein A site: PBM
Protein A modification: (TPO)147
Protein A sequence: SRSRRRETAL
Protein A construct: Biotin-ttds-SRSRRREpTAL
Protein B protein: DLG4
Protein B fragment: 306-403
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.03
Normalized holdup BI: 0.03
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
PUBMED ID of publication: 36115835
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 4
Protein A protein: HPV33-E6
Protein A fragment: 140-149
Protein A site: PBM
Protein A sequence: SRSRRRETAL
Protein A construct: biotin-ttds-SRSRRRETAL
Protein B protein: DLG4
Protein B fragment: 306-403
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.68
Holdup BI standard deviation: 0.05
Normalized holdup BI: 0.78
Normalized holdup BI standard deviation: 0.06
Immobilized partner concentration in holdup experiment (10-6M): 20.2
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