Details of experiments between the fragments P27228~140-149 and Q14160~999-1093
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: HPV35-E6
Protein A fragment: 140-149
Protein A site: PBM
Protein A sequence: SKPTRRETEV
Protein A construct: biotin-ttds-SKPTRRETEV
Protein B protein: SCRIB
Protein B fragment: 999-1093
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.78
Immobilized partner concentration in holdup experiment (10-6M): 39.7
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
Experiment series 2
Protein A protein: HPV35-E6
Protein A fragment: 140-149
Protein A site: PBM
Protein A sequence: SKPTRRETEV
Protein A construct: biotin-ttds-SKPTRRETEV
Protein B protein: SCRIB
Protein B fragment: 999-1093
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ
Measured dissociation constant (10-6M): 9.62
Standard deviation of dissociation constant (10-6M): 0.42
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 5.02
Experiment series 3
Protein A protein: HPV35-E6
Protein A fragment: 140-149
Protein A site: PBM
Protein A modification: (TPO)147
Protein A sequence: SKPTRRETEV
Protein A construct: Biotin-ttds-SKPTRREpTEV
Protein B protein: SCRIB
Protein B fragment: 999-1093
Protein B site: PDZ_3
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: HPV35-E6
Protein A fragment: 140-149
Protein A site: PBM
Protein A sequence: SKPTRRETEV
Protein A construct: biotin-ttds-SKPTRRETEV
Protein B protein: SCRIB
Protein B fragment: 999-1093
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.5
Holdup BI standard deviation: 0.02
Normalized holdup BI: 0.56
Normalized holdup BI standard deviation: 0.02
Immobilized partner concentration in holdup experiment (10-6M): 18.55
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: 4.89
Experiment series 5
Protein A protein: HPV35-E6
Protein A fragment: 140-149
Protein A site: PBM
Protein A sequence: SKPTRRETEV
Protein A construct: biotin-ttds-SKPTRRETEV
Protein B protein: SCRIB
Protein B fragment: 999-1093
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.33
Normalized holdup BI: 0.4
Immobilized partner concentration in holdup experiment (10-6M): 18.55
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: 4.6
Experiment series 6
Protein A protein: HPV35-E6
Protein A fragment: 140-149
Protein A site: PBM
Protein A sequence: SKPTRRETEV
Protein A construct: biotin-ttds-SKPTRRETEV
Protein B protein: SCRIB
Protein B fragment: 999-1093
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.5
Normalized holdup BI: 0.59
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: 4.97