Details of experiments between Q07157 and P50479
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: PDLIM4
Protein B fragment: 237-246
Protein B site: PBM
Protein B modification: iso2
Protein B sequence: VYPNAKVELV
Protein B construct: biotin-ado-ado-VYPNAKVELV
Average holdup BI: 0.04
Normalized holdup BI: 0.06
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: Crude peptide synthesis product. The peptide concentration in the affinity conversion step may differ from reality causing a systematic error in reported affinities.
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 3.52
Experiment series 2
Protein A protein: TJP1
Protein A fragment: 415-516
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: PDLIM4
Protein B fragment: 237-246
Protein B site: PBM
Protein B modification: iso2
Protein B sequence: VYPNAKVELV
Protein B construct: biotin-ado-ado-VYPNAKVELV
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
Experimental details: Crude peptide synthesis product. The peptide concentration in the affinity conversion step may differ from reality causing a systematic error in reported affinities.
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: TJP1
Protein A fragment: 184-264
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: PDLIM4
Protein B fragment: 321-330
Protein B site: PBM
Protein B sequence: PSASCAVTAA
Protein B construct: Biotin-Ahx-PSASCAVTAA
Average holdup BI: -0.04
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 4
Protein A protein: TJP1
Protein A fragment: 415-516
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: PDLIM4
Protein B fragment: 321-330
Protein B site: PBM
Protein B sequence: PSASCAVTAA
Protein B construct: Biotin-Ahx-PSASCAVTAA
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.