Details of experiments between Q8NI35 and P11274
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: Inadl
Protein A fragment: 359-469
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: BCR
Protein B fragment: 1262-1271
Protein B site: PBM
Protein B sequence: RQSILFSTEV
Protein B construct: biotin-ado-ado-RQSILFSTEV
Average holdup BI: 0.42
Normalized holdup BI: 0.42
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: 4.66
Experiment series 2
Protein A protein: Inadl
Protein A fragment: 1425-1530
Protein A site: PDZ_8
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: BCR
Protein B fragment: 1262-1271
Protein B site: PBM
Protein B sequence: RQSILFSTEV
Protein B construct: biotin-ado-ado-RQSILFSTEV
Average holdup BI: 0.32
Normalized holdup BI: 0.34
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: 4.49
Experiment series 3
Protein A protein: Inadl
Protein A fragment: 1673-1763
Protein A site: PDZ_10
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: BCR
Protein B fragment: 1262-1271
Protein B site: PBM
Protein B sequence: RQSILFSTEV
Protein B construct: biotin-ado-ado-RQSILFSTEV
Average holdup BI: 0.26
Normalized holdup BI: 0.26
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: 4.32
Experiment series 4
Protein A protein: Inadl
Protein A fragment: 1530-1615
Protein A site: PDZ_9
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: BCR
Protein B fragment: 1262-1271
Protein B site: PBM
Protein B sequence: RQSILFSTEV
Protein B construct: biotin-ado-ado-RQSILFSTEV
Average holdup BI: 0.18
Normalized holdup BI: 0.18
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: 4.09
Experiment series 5
Protein A protein: Inadl
Protein A fragment: 1058-1167
Protein A site: PDZ_6
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: BCR
Protein B fragment: 1262-1271
Protein B site: PBM
Protein B sequence: RQSILFSTEV
Protein B construct: biotin-ado-ado-RQSILFSTEV
Average holdup BI: 0.13
Normalized holdup BI: 0.13
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: 3.93
Experiment series 6
Protein A protein: Inadl
Protein A fragment: 113-229
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: BCR
Protein B fragment: 1262-1271
Protein B site: PBM
Protein B sequence: RQSILFSTEV
Protein B construct: biotin-ado-ado-RQSILFSTEV
Average holdup BI: 0.12
Normalized holdup BI: 0.12
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: 3.88
Experiment series 7
Protein A protein: Inadl
Protein A fragment: 673-780
Protein A site: PDZ_5
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: BCR
Protein B fragment: 1262-1271
Protein B site: PBM
Protein B sequence: RQSILFSTEV
Protein B construct: biotin-ado-ado-RQSILFSTEV
Average holdup BI: 0.05
Normalized holdup BI: 0.05
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: 3.49
Experiment series 8
Protein A protein: Inadl
Protein A fragment: 238-335
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: BCR
Protein B fragment: 1262-1271
Protein B site: PBM
Protein B sequence: RQSILFSTEV
Protein B construct: biotin-ado-ado-RQSILFSTEV
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
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 9
Protein A protein: Inadl
Protein A fragment: 547-651
Protein A site: PDZ_4
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: BCR
Protein B fragment: 1262-1271
Protein B site: PBM
Protein B sequence: RQSILFSTEV
Protein B construct: biotin-ado-ado-RQSILFSTEV
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
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.