Details of experiments between Q07157 and P0DTC4
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: TJP1
Protein A fragment: 184-264
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: SARS-CoV2-E
Protein B fragment: 66-75
Protein B site: PBM
Protein B sequence: NSSRVPDLLV
Protein B construct: biotin-NLNSSRVPDLLV
Average holdup BI: 0.87
Holdup BI standard deviation: 0
Immobilized partner concentration in holdup experiment (10-6M): 20
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: 5.61
Experiment series 2
Protein A protein: TJP1
Protein A fragment: 14-113
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: SARS-CoV2-E
Protein B fragment: 66-75
Protein B site: PBM
Protein B sequence: NSSRVPDLLV
Protein B construct: biotin-NLNSSRVPDLLV
Average holdup BI: -0.11
Holdup BI standard deviation: 0.1
Immobilized partner concentration in holdup experiment (10-6M): 20
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
The affinity was below the detection threshold of the assay.
Experiment series 3
Protein A protein: TJP1
Protein A fragment: 415-516
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: SARS-CoV2-E
Protein B fragment: 66-75
Protein B site: PBM
Protein B sequence: NSSRVPDLLV
Protein B construct: biotin-NLNSSRVPDLLV
Average holdup BI: -0.07
Immobilized partner concentration in holdup experiment (10-6M): 20
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
The affinity was below the detection threshold of the assay.
Experiment series 4
Protein A protein: TJP1
Protein A fragment: 184-264
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: SARS-CoV2-E
Protein B fragment: 66-75
Protein B site: PBM
Protein B sequence: NSSRVPDLLV
Protein B construct: biotin-NLNSSRVPDLLV
Average holdup BI: 0.69
Normalized holdup BI: 0.83
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
Measured pKd: 5.52
Experiment series 5
Protein A protein: TJP1
Protein A fragment: 415-516
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: SARS-CoV2-E
Protein B fragment: 66-75
Protein B site: PBM
Protein B sequence: NSSRVPDLLV
Protein B construct: biotin-NLNSSRVPDLLV
Average holdup BI: 0.03
Normalized holdup BI: 0.03
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.