Details of experiments between the fragments Q14160~999-1093 and P06463~149-158
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: SCRIB
Protein A fragment: 999-1093
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: HPV18-E6
Protein B fragment: 149-158
Protein B site: PBM
Protein B sequence: RLQRRRETQV
Protein B construct: biotin-ttds-RLQRRRETQV
Average holdup BI: 0.37
Holdup BI standard deviation: 0.02
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: 4.7
Experiment series 2
Protein A protein: SCRIB
Protein A fragment: 999-1093
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: HPV18-E6
Protein B fragment: 149-158
Protein B site: PBM
Protein B sequence: RLQRRRETQV
Protein B construct: biotin-ttds-RLQRRRETQV
Measured dissociation constant (10-6M): 14.73
Standard deviation of dissociation constant (10-6M): 0.82
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 4.83
Experiment series 3
Protein A protein: SCRIB
Protein A fragment: 999-1093
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: HPV18-E6
Protein B fragment: 149-158
Protein B site: PBM
Protein B modification: (TPO)156
Protein B sequence: RLQRRRETQV
Protein B construct: Biotin-ttds-RLQRRREpTQV
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: SCRIB
Protein A fragment: 999-1093
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: HPV18-E6
Protein B fragment: 149-158
Protein B site: PBM
Protein B sequence: RLQRRRETQV
Protein B construct: biotin-ttds-RLQRRRETQV
Average holdup BI: 0.66
Holdup BI standard deviation: 0.11
Normalized holdup BI: 0.74
Normalized holdup BI standard deviation: 0.12
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: 4.84
Experiment series 5
Protein A protein: SCRIB
Protein A fragment: 999-1093
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: HPV18-E6
Protein B fragment: 149-158
Protein B site: PBM
Protein B sequence: RLQRRRETQV
Protein B construct: biotin-ttds-RLQRRRETQV
Average holdup BI: 0.41
Normalized holdup BI: 0.49
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: 4.37
Experiment series 6
Protein A protein: SCRIB
Protein A fragment: 999-1093
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: HPV18-E6
Protein B fragment: 149-158
Protein B site: PBM
Protein B sequence: RLQRRRETQV
Protein B construct: biotin-ttds-RLQRRRETQV
Average holdup BI: 0.49
Normalized holdup BI: 0.58
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: 4.94