Details of experiments between the fragments P03409~344-353 and Q14160~718-814
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: HTLV1-TAX1
Protein A fragment: 344-353
Protein A site: PBM
Protein A sequence: SEKHFRETEV
Protein A construct: biotin-ttds-SEKHFRETEV
Protein B protein: SCRIB
Protein B fragment: 718-814
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.91
Holdup BI standard deviation: 0.01
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: 5.81
Experiment series 2
Protein A protein: HTLV1-TAX1
Protein A fragment: 344-353
Protein A site: PBM
Protein A sequence: SEKHFRETEV
Protein A construct: biotin-ttds-SEKHFRETEV
Protein B protein: SCRIB
Protein B fragment: 718-814
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Measured dissociation constant (10-6M): 2.51
Standard deviation of dissociation constant (10-6M): 0.08
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 5.6
Experiment series 3
Protein A protein: HTLV1-TAX1
Protein A fragment: 344-353
Protein A site: PBM
Protein A sequence: SEKHFRETEV
Protein A construct: biotin-ttds-SEKHFRETEV
Protein B protein: SCRIB
Protein B fragment: 718-814
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.79
Holdup BI standard deviation: 0.04
Normalized holdup BI: 0.83
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: 7
Measured pKd: 5.63
Experiment series 4
Protein A protein: HTLV1-TAX1
Protein A fragment: 344-353
Protein A site: PBM
Protein A sequence: SEKHFRETEV
Protein A construct: Biotin-ado-ado-SEKHFRETEV
Protein B protein: SCRIB
Protein B fragment: 718-814
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.87
Normalized holdup BI: 0.87
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: 5.67