Details of experiments between the fragments Q9NYB5~703-712 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: SLCO1C1
Protein A fragment: 703-712
Protein A site: PBM
Protein A sequence: NYWPGKETQL
Protein A construct: biotin-ttds-NYWPGKETQL
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): 11.98
Standard deviation of dissociation constant (10-6M): 0.8
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 4.92
Experiment series 2
Protein A protein: SLCO1C1
Protein A fragment: 703-712
Protein A site: PBM
Protein A sequence: NYWPGKETQL
Protein A construct: biotin-ttds-NYWPGKETQL
Protein B protein: SCRIB
Protein B fragment: 718-814
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.42
Immobilized partner concentration in holdup experiment (10-6M): 5.5
Experiment method: SAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normal layout
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 5.29
Experiment series 3
Protein A protein: SLCO1C1
Protein A fragment: 703-712
Protein A site: PBM
Protein A sequence: NYWPGKETQL
Protein A construct: biotin-ttds-NYWPGKETQL
Protein B protein: SCRIB
Protein B fragment: 718-814
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.57
Holdup BI standard deviation: 0.05
Normalized holdup BI: 0.6
Normalized holdup BI standard deviation: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 11.3
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: 5
Measured pKd: 5.23
Experiment series 4
Protein A protein: SLCO1C1
Protein A fragment: 703-712
Protein A site: PBM
Protein A sequence: NYWPGKETQL
Protein A construct: biotin-ttds-NYWPGKETQL
Protein B protein: SCRIB
Protein B fragment: 718-814
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.41
Normalized holdup BI: 0.51
Immobilized partner concentration in holdup experiment (10-6M): 11.3
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
Experimental details: not reported internal standards of holdup layout #5
Number of measurements: 1
Measured pKd: 5.05
Experiment series 5
Protein A protein: SLCO1C1
Protein A fragment: 703-712
Protein A site: PBM
Protein A sequence: NYWPGKETQL
Protein A construct: biotin-ttds-NYWPGKETQL
Protein B protein: SCRIB
Protein B fragment: 718-814
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.46
Normalized holdup BI: 0.5
Immobilized partner concentration in holdup experiment (10-6M): 11.3
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
Experimental details: not reported internal standards of holdup layout #5
Number of measurements: 1
Measured pKd: 5.03
Experiment series 6
Protein A protein: SLCO1C1
Protein A fragment: 703-712
Protein A site: PBM
Protein A sequence: NYWPGKETQL
Protein A construct: Biotin-ado-ado-NYWPGKETQL
Protein B protein: SCRIB
Protein B fragment: 718-814
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.71
Normalized holdup BI: 0.71
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: 5.2