Details of experiments between the fragments P27228~140-149 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: 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: 718-814
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.86
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: 5.24
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: 718-814
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Measured dissociation constant (10-6M): 17.97
Standard deviation of dissociation constant (10-6M): 1.01
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 4.75
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: 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: 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: 718-814
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.36
Holdup BI standard deviation: 0.04
Normalized holdup BI: 0.38
Normalized holdup BI standard deviation: 0.04
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: 8
Measured pKd: 4.56
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: 718-814
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.24
Normalized holdup BI: 0.3
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: 4.4
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: 718-814
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.32
Normalized holdup BI: 0.35
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: 4.5
Experiment series 7
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: 718-814
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.5
Normalized holdup BI: 0.5
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: 4.8