Details of experiments between the fragments Q12959~221-314 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: DLG1
Protein A fragment: 221-314
Protein A site: PDZ_1
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: 0.27
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: 4.8
Experiment series 2
Protein A protein: DLG1
Protein A fragment: 221-314
Protein A site: PDZ_1
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: -0.02
Normalized holdup BI: -0.02
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: DLG1
Protein A fragment: 221-314
Protein A site: PDZ_1
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: 0.31
Holdup BI standard deviation: 0.06
Normalized holdup BI: 0.37
Normalized holdup BI standard deviation: 0.07
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: 4.5