Details of experiments between the fragments P06463~149-158 and Q86UL8~410-522
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: MAGI2
Protein B fragment: 410-522
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.86
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: 1
Measured pKd: 5.79
Experiment series 2
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: MAGI2
Protein B fragment: 410-522
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ
Measured dissociation constant (10-6M): 1.03
Standard deviation of dissociation constant (10-6M): 0.03
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 5.99
Experiment series 3
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: MAGI2
Protein B fragment: 410-522
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.02
Normalized holdup BI: 0.02
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: HPV18-E6
Protein A fragment: 149-158
Protein A site: PBM
Protein A sequence: RLQRRRETQV
Protein A construct: biotin-ttds-RLQRRRETQV
Protein B protein: MAGI2
Protein B fragment: 410-522
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.77
Normalized holdup BI: 0.85
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: 1
Measured pKd: 5.16
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: MAGI2
Protein B fragment: 410-522
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.77
Normalized holdup BI: 0.92
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: 5.5
Experiment series 6
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: MAGI2
Protein B fragment: 410-522
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.85
Normalized holdup BI: 0.93
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: 5.53
Experiment series 7
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: MAGI2
Protein B fragment: 410-522
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.84
Normalized holdup BI: 0.93
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: 5.96