Details of experiments between the fragments Q5T5U3~47-159 and P03126~149-158
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: ARHGAP21
Protein A fragment: 47-159
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: HPV16-E6
Protein B fragment: 149-158
Protein B site: PBM
Protein B modification: (TPO)156
Protein B sequence: SSRTRRETQL
Protein B construct: Biotin-ttds-SSRTRREpTQL
Average holdup BI: 0.2
Immobilized partner concentration in holdup experiment (10-6M): 18
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: 4.17
Experiment series 2
Protein A protein: ARHGAP21
Protein A fragment: 47-159
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: HPV16-E6
Protein B fragment: 149-158
Protein B site: PBM
Protein B sequence: SSRTRRETQL
Protein B construct: biotin-ttds-SSRTRRETQL
Average holdup BI: 0
Holdup BI standard deviation: 0.08
Immobilized partner concentration in holdup experiment (10-6M): 15
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: 6
The affinity was below the detection threshold of the assay.
Experiment series 3
Protein A protein: ARHGAP21
Protein A fragment: 47-159
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: HPV16-E6
Protein B fragment: 149-158
Protein B site: PBM
Protein B modification: (TPO)156
Protein B sequence: SSRTRRETQL
Protein B construct: Biotin-ttds-SSRTRREpTQL
Average holdup BI: 0.16
Normalized holdup BI: 0.16
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
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 4.05
Experiment series 4
Protein A protein: ARHGAP21
Protein A fragment: 47-159
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: HPV16-E6
Protein B fragment: 149-158
Protein B site: PBM
Protein B sequence: SSRTRRETQL
Protein B construct: biotin-ttds-SSRTRRETQL
Average holdup BI: 0.03
Holdup BI standard deviation: 0
Normalized holdup BI: 0.03
Normalized holdup BI standard deviation: 0
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
PUBMED ID of publication: 36115835
Number of measurements: 2
The affinity was below the detection threshold of the assay.
Experiment series 5
Protein A protein: ARHGAP21
Protein A fragment: 47-159
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: HPV16-E6
Protein B fragment: 149-158
Protein B site: PBM
Protein B sequence: SSRTRRETQL
Protein B construct: biotin-ttds-SSRTRRETQL
Average holdup BI: 0.05
Normalized holdup BI: 0.05
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: not reported internal standards of holdup layout #5
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 6
Protein A protein: ARHGAP21
Protein A fragment: 47-159
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: HPV16-E6
Protein B fragment: 149-158
Protein B site: PBM
Protein B sequence: SSRTRRETQL
Protein B construct: Biotin-ado-ado-SSRTRRETQL
Average holdup BI: 0.13
Normalized holdup BI: 0.13
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: 3.92