Details of experiments between the fragments P06427~140-149 and Q9NZN5~67-151
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: HPV33-E6
Protein A fragment: 140-149
Protein A site: PBM
Protein A sequence: SRSRRRETAL
Protein A construct: biotin-ttds-SRSRRRETAL
Protein B protein: ARHGEF12
Protein B fragment: 67-151
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.37
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: 5.03
Experiment series 2
Protein A protein: HPV33-E6
Protein A fragment: 140-149
Protein A site: PBM
Protein A modification: (TPO)147
Protein A sequence: SRSRRRETAL
Protein A construct: Biotin-ttds-SRSRRREpTAL
Protein B protein: ARHGEF12
Protein B fragment: 67-151
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: -0.03
Normalized holdup BI: -0.04
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: HPV33-E6
Protein A fragment: 140-149
Protein A site: PBM
Protein A sequence: SRSRRRETAL
Protein A construct: biotin-ttds-SRSRRRETAL
Protein B protein: ARHGEF12
Protein B fragment: 67-151
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.47
Holdup BI standard deviation: 0.05
Normalized holdup BI: 0.52
Normalized holdup BI standard deviation: 0.05
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: 4.78