Details of experiments between the fragments Q13884~107-196 and P03409~344-353
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: SNTB1
Protein A fragment: 107-196
Protein A site: PDZ
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: 0.93
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: 1
Measured pKd: 5.91
Experiment series 2
Protein A protein: SNTB1
Protein A fragment: 107-196
Protein A site: PDZ
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): 0.93
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: 6.03
Experiment series 3
Protein A protein: SNTB1
Protein A fragment: 107-196
Protein A site: PDZ
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: 0.78
Normalized holdup BI: 0.94
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: 6.12
Experiment series 4
Protein A protein: SNTB1
Protein A fragment: 107-196
Protein A site: PDZ
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: 0.69
Normalized holdup BI: 0.77
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: 5.35