Details of experiments between the fragments P03409~344-353 and Q86UL8~410-522
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: MAGI2
Protein B fragment: 410-522
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.6
Holdup BI standard deviation: 0.02
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: 4.96
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: MAGI2
Protein B fragment: 410-522
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ
Measured dissociation constant (10-6M): 16.68
Standard deviation of dissociation constant (10-6M): 0.97
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 4.78
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: MAGI2
Protein B fragment: 410-522
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.31
Normalized holdup BI: 0.34
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
Measured pKd: 4.57
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: MAGI2
Protein B fragment: 410-522
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.46
Normalized holdup BI: 0.51
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: 4.81