Details of experiments between the fragments P06463~149-158 and Q14160~718-814
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: SCRIB
Protein B fragment: 718-814
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.14
Holdup BI standard deviation: 0.09
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: 4.1
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: SCRIB
Protein B fragment: 718-814
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Measured dissociation constant (10-6M): 31.99
Standard deviation of dissociation constant (10-6M): 7.98
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 4.49
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: SCRIB
Protein B fragment: 718-814
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: -0.01
Normalized holdup BI: -0.01
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: SCRIB
Protein B fragment: 718-814
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.41
Holdup BI standard deviation: 0.04
Normalized holdup BI: 0.43
Normalized holdup BI standard deviation: 0.04
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: 7
Measured pKd: 4.26
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: SCRIB
Protein B fragment: 718-814
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.34
Normalized holdup BI: 0.43
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: 4.25
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: SCRIB
Protein B fragment: 718-814
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.38
Normalized holdup BI: 0.42
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: 4.24
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: SCRIB
Protein B fragment: 718-814
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.54
Normalized holdup BI: 0.54
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: 4.87