Details of experiments between the fragments P06463~149-158 and P78352~62-152
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: HPV18-E6
Protein A fragment: 149-158
Protein A site: PBM
Protein A sequence: RLQRRRETQV
Protein A construct: biotin-ttds-RLQRRRETQV
Protein B protein: DLG4
Protein B fragment: 62-152
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.63
Holdup BI standard deviation: 0.07
Immobilized partner concentration in holdup experiment (10-6M): 13.3
Experiment method: CALIP_holdup
Details of affinity fitting: BSA and lysozyme internal standards + bacterial cell lysate + normal layout
PUBMED ID of publication: 36115835
Number of measurements: 2
Measured pKd: 5.2
Experiment series 2
Protein A protein: HPV18-E6
Protein A fragment: 149-158
Protein A site: PBM
Protein A modification: (TPO)156
Protein A sequence: RLQRRRETQV
Protein A construct: Biotin-ttds-RLQRRREpTQV
Protein B protein: DLG4
Protein B fragment: 62-152
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: -0.07
Normalized holdup BI: -0.09
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 3
Protein A protein: HPV18-E6
Protein A fragment: 149-158
Protein A site: PBM
Protein A sequence: RLQRRRETQV
Protein A construct: biotin-ttds-RLQRRRETQV
Protein B protein: DLG4
Protein B fragment: 62-152
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.7
Holdup BI standard deviation: 0
Normalized holdup BI: 0.83
Normalized holdup BI standard deviation: 0
Immobilized partner concentration in holdup experiment (10-6M): 43
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: 5.11
Experiment series 4
Protein A protein: HPV18-E6
Protein A fragment: 149-158
Protein A site: PBM
Protein A sequence: RLQRRRETQV
Protein A construct: biotin-ttds-RLQRRRETQV
Protein B protein: DLG4
Protein B fragment: 62-152
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.67
Normalized holdup BI: 0.67
Immobilized partner concentration in holdup experiment (10-6M): 43
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 #5
Number of measurements: 1
Measured pKd: 4.7
Experiment series 5
Protein A protein: HPV18-E6
Protein A fragment: 149-158
Protein A site: PBM
Protein A sequence: RLQRRRETQV
Protein A construct: biotin-ttds-RLQRRRETQV
Protein B protein: DLG4
Protein B fragment: 62-152
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.68
Normalized holdup BI: 0.8
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: 5.45