Details of experiments between the fragments P03126~149-158 and Q96QZ7~456-580
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: HPV16-E6
Protein A fragment: 149-158
Protein A site: PBM
Protein A modification: (TPO)156
Protein A sequence: SSRTRRETQL
Protein A construct: Biotin-ttds-SSRTRREpTQL
Protein B protein: MAGI1
Protein B fragment: 456-580
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: -0.03
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
The affinity was below the detection threshold of the assay.
Experiment series 2
Protein A protein: HPV16-E6
Protein A fragment: 149-158
Protein A site: PBM
Protein A sequence: SSRTRRETQL
Protein A construct: biotin-ttds-SSRTRRETQL
Protein B protein: MAGI1
Protein B fragment: 456-580
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.87
Holdup BI standard deviation: 0.09
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: 5
Measured pKd: 5.74
Experiment series 3
Protein A protein: HPV16-E6
Protein A fragment: 149-158
Protein A site: PBM
Protein A sequence: SSRTRRETQL
Protein A construct: biotin-ttds-SSRTRRETQL
Protein B protein: MAGI1
Protein B fragment: 456-580
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ
Measured dissociation constant (10-6M): 4.1
Standard deviation of dissociation constant (10-6M): 0.12
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 5.39
Experiment series 4
Protein A protein: HPV16-E6
Protein A fragment: 149-158
Protein A site: PBM
Protein A modification: (TPO)156
Protein A sequence: SSRTRRETQL
Protein A construct: Biotin-ttds-SSRTRREpTQL
Protein B protein: MAGI1
Protein B fragment: 456-580
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ
Measured dissociation constant (10-6M): 2520
Standard deviation of dissociation constant (10-6M): 350
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 2.6
Experiment series 5
Protein A protein: HPV16-E6
Protein A fragment: 149-158
Protein A site: PBM
Protein A modification: (TPO)156
Protein A sequence: SSRTRRETQL
Protein A construct: Biotin-ttds-SSRTRREpTQL
Protein B protein: MAGI1
Protein B fragment: 456-580
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: -0.02
Normalized holdup BI: -0.02
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
The affinity was below the detection threshold of the assay.
Experiment series 6
Protein A protein: HPV16-E6
Protein A fragment: 149-158
Protein A site: PBM
Protein A sequence: SSRTRRETQL
Protein A construct: biotin-ttds-SSRTRRETQL
Protein B protein: MAGI1
Protein B fragment: 456-580
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.81
Holdup BI standard deviation: 0.03
Normalized holdup BI: 0.92
Normalized holdup BI standard deviation: 0.04
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
Measured pKd: 5.91
Experiment series 7
Protein A protein: HPV16-E6
Protein A fragment: 149-158
Protein A site: PBM
Protein A sequence: SSRTRRETQL
Protein A construct: biotin-ttds-SSRTRRETQL
Protein B protein: MAGI1
Protein B fragment: 456-580
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.75
Normalized holdup BI: 0.9
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
Measured pKd: 5.78
Experiment series 8
Protein A protein: HPV16-E6
Protein A fragment: 149-158
Protein A site: PBM
Protein A sequence: SSRTRRETQL
Protein A construct: Biotin-ado-ado-SSRTRRETQL
Protein B protein: MAGI1
Protein B fragment: 456-580
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.84
Normalized holdup BI: 0.93
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: 5.98
Experiment series 9
Protein A protein: HPV16-E6
Protein A fragment: 149-158
Protein A site: PBM
Protein A sequence: SSRTRRETQL
Protein A construct: Biotin-ttds-SSRTRRETQL
Protein B protein: MAGI1
Protein B fragment: 456-580
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ
Measured dissociation constant (10-6M): 3.4
Standard deviation of dissociation constant (10-6M): 0.8
Experiment method: SPR
PUBMED ID of publication: 30726710
Number of measurements: 1
Measured pKd: 5.47