Details of experiments between the fragments O14907~7-124 and P06427~140-149
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: TX1BP3
Protein A fragment: 7-124
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: HPV33-E6
Protein B fragment: 140-149
Protein B site: PBM
Protein B sequence: SRSRRRETAL
Protein B construct: biotin-ttds-SRSRRRETAL
Average holdup BI: 0.47
Immobilized partner concentration in holdup experiment (10-6M): 7
Experiment method: SAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normal layout
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 5.24
Experiment series 2
Protein A protein: TX1BP3
Protein A fragment: 7-124
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: HPV33-E6
Protein B fragment: 140-149
Protein B site: PBM
Protein B modification: (TPO)147
Protein B sequence: SRSRRRETAL
Protein B construct: Biotin-ttds-SRSRRREpTAL
Average holdup BI: 0
Normalized holdup BI: 0
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 3
Protein A protein: TX1BP3
Protein A fragment: 7-124
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: HPV33-E6
Protein B fragment: 140-149
Protein B site: PBM
Protein B sequence: SRSRRRETAL
Protein B construct: biotin-ttds-SRSRRRETAL
Average holdup BI: 0.61
Holdup BI standard deviation: 0.02
Normalized holdup BI: 0.67
Normalized holdup BI standard deviation: 0.02
Immobilized partner concentration in holdup experiment (10-6M): 20.2
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: 5.07