Details of experiments between the fragments P06463~149-158 and Q12959~457-570
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: DLG1
Protein B fragment: 457-570
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.61
Holdup BI standard deviation: 0.03
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: 5.16
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: DLG1
Protein B fragment: 457-570
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: -0.03
Normalized holdup BI: -0.04
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: DLG1
Protein B fragment: 457-570
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.67
Holdup BI standard deviation: 0.05
Normalized holdup BI: 0.81
Normalized holdup BI standard deviation: 0.06
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: 5.02
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: DLG1
Protein B fragment: 457-570
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.58
Normalized holdup BI: 0.75
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: 4.88
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: DLG1
Protein B fragment: 457-570
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.52
Normalized holdup BI: 0.78
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: 5.37