Details of experiments between Q5T2W1 and P03409
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: NHERF3
Protein A fragment: 2-107
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: HTLV1-TAX1
Protein B fragment: 344-353
Protein B site: PBM
Protein B sequence: SEKHFRETEV
Protein B construct: biotin-ttds-SEKHFRETEV
Average holdup BI: 0.64
Holdup BI standard deviation: 0.04
Immobilized partner concentration in holdup experiment (10-6M): 19
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: 5.03
Experiment series 2
Protein A protein: NHERF3
Protein A fragment: 132-224
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: HTLV1-TAX1
Protein B fragment: 344-353
Protein B site: PBM
Protein B sequence: SEKHFRETEV
Protein B construct: biotin-ttds-SEKHFRETEV
Average holdup BI: 0.06
Holdup BI standard deviation: 0.11
Immobilized partner concentration in holdup experiment (10-6M): 19
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: NHERF3
Protein A fragment: 238-329
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: HTLV1-TAX1
Protein B fragment: 344-353
Protein B site: PBM
Protein B sequence: SEKHFRETEV
Protein B construct: biotin-ttds-SEKHFRETEV
Average holdup BI: -0.04
Holdup BI standard deviation: 0.06
Immobilized partner concentration in holdup experiment (10-6M): 19
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: 4
The affinity was below the detection threshold of the assay.
Experiment series 4
Protein A protein: NHERF3
Protein A fragment: 375-459
Protein A site: PDZ_4
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: HTLV1-TAX1
Protein B fragment: 344-353
Protein B site: PBM
Protein B sequence: SEKHFRETEV
Protein B construct: biotin-ttds-SEKHFRETEV
Average holdup BI: 0.04
Holdup BI standard deviation: 0.04
Immobilized partner concentration in holdup experiment (10-6M): 19
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 5
Protein A protein: NHERF3
Protein A fragment: 2-107
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: HTLV1-TAX1
Protein B fragment: 344-353
Protein B site: PBM
Protein B sequence: SEKHFRETEV
Protein B construct: biotin-ttds-SEKHFRETEV
Measured dissociation constant (10-6M): 25.08
Standard deviation of dissociation constant (10-6M): 2.03
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 4.6
Experiment series 6
Protein A protein: NHERF3
Protein A fragment: 2-107
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: HTLV1-TAX1
Protein B fragment: 344-353
Protein B site: PBM
Protein B sequence: SEKHFRETEV
Protein B construct: biotin-ttds-SEKHFRETEV
Average holdup BI: 0.38
Holdup BI standard deviation: 0.04
Normalized holdup BI: 0.41
Normalized holdup BI standard deviation: 0.04
Immobilized partner concentration in holdup experiment (10-6M): 15
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: 4.71
Experiment series 7
Protein A protein: NHERF3
Protein A fragment: 238-329
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: HTLV1-TAX1
Protein B fragment: 344-353
Protein B site: PBM
Protein B sequence: SEKHFRETEV
Protein B construct: biotin-ttds-SEKHFRETEV
Average holdup BI: 0.03
Normalized holdup BI: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 15
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 8
Protein A protein: NHERF3
Protein A fragment: 375-459
Protein A site: PDZ_4
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: HTLV1-TAX1
Protein B fragment: 344-353
Protein B site: PBM
Protein B sequence: SEKHFRETEV
Protein B construct: biotin-ttds-SEKHFRETEV
Average holdup BI: 0.03
Normalized holdup BI: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 15
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 9
Protein A protein: NHERF3
Protein A fragment: 2-107
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: HTLV1-TAX1
Protein B fragment: 344-353
Protein B site: PBM
Protein B sequence: SEKHFRETEV
Protein B construct: Biotin-ado-ado-SEKHFRETEV
Average holdup BI: 0.47
Normalized holdup BI: 0.5
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. Not reported internal standards of holdup layout #6
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 4.8
Experiment series 10
Protein A protein: NHERF3
Protein A fragment: 375-459
Protein A site: PDZ_4
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: HTLV1-TAX1
Protein B fragment: 344-353
Protein B site: PBM
Protein B sequence: SEKHFRETEV
Protein B construct: Biotin-ado-ado-SEKHFRETEV
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. Not reported internal standards of holdup layout #6
PUBMED ID of publication: 36115835
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 11
Protein A protein: NHERF3
Protein A fragment: 238-329
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: HTLV1-TAX1
Protein B fragment: 344-353
Protein B site: PBM
Protein B sequence: SEKHFRETEV
Protein B construct: Biotin-ado-ado-SEKHFRETEV
Average holdup BI: -0.07
Normalized holdup BI: -0.07
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. Not reported internal standards of holdup layout #6
PUBMED ID of publication: 36115835
Number of measurements: 1
The affinity was below the detection threshold of the assay.