Details of experiments between the fragments Q14160~858-949 and Q9NYB5~703-712
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: SCRIB
Protein A fragment: 858-949
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: SLCO1C1
Protein B fragment: 703-712
Protein B site: PBM
Protein B sequence: NYWPGKETQL
Protein B construct: biotin-ttds-NYWPGKETQL
Average holdup BI: -0.05
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
The affinity was below the detection threshold of the assay.
Experiment series 2
Protein A protein: SCRIB
Protein A fragment: 858-949
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: SLCO1C1
Protein B fragment: 703-712
Protein B site: PBM
Protein B sequence: NYWPGKETQL
Protein B construct: biotin-ttds-NYWPGKETQL
Average holdup BI: 0.12
Holdup BI standard deviation: 0.05
Normalized holdup BI: 0.16
Normalized holdup BI standard deviation: 0.06
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: 3
Measured pKd: 4.24
Experiment series 3
Protein A protein: SCRIB
Protein A fragment: 858-949
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: SLCO1C1
Protein B fragment: 703-712
Protein B site: PBM
Protein B sequence: NYWPGKETQL
Protein B construct: biotin-ttds-NYWPGKETQL
Average holdup BI: 0.12
Normalized holdup BI: 0.16
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: 4.25
Experiment series 4
Protein A protein: SCRIB
Protein A fragment: 858-949
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: SLCO1C1
Protein B fragment: 703-712
Protein B site: PBM
Protein B sequence: NYWPGKETQL
Protein B construct: Biotin-ado-ado-NYWPGKETQL
Average holdup BI: 0.25
Normalized holdup BI: 0.25
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: 4.28