Details of experiments between the fragments Q12959~221-314 and P27228~140-149
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: DLG1
Protein A fragment: 221-314
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: HPV35-E6
Protein B fragment: 140-149
Protein B site: PBM
Protein B sequence: SKPTRRETEV
Protein B construct: biotin-ttds-SKPTRRETEV
Average holdup BI: 0.74
Immobilized partner concentration in holdup experiment (10-6M): 39.7
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: 4.88
Experiment series 2
Protein A protein: DLG1
Protein A fragment: 221-314
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: HPV35-E6
Protein B fragment: 140-149
Protein B site: PBM
Protein B modification: (TPO)147
Protein B sequence: SKPTRRETEV
Protein B construct: Biotin-ttds-SKPTRREpTEV
Average holdup BI: -0.03
Normalized holdup BI: -0.03
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 3
Protein A protein: DLG1
Protein A fragment: 221-314
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: HPV35-E6
Protein B fragment: 140-149
Protein B site: PBM
Protein B sequence: SKPTRRETEV
Protein B construct: biotin-ttds-SKPTRRETEV
Average holdup BI: 0.53
Holdup BI standard deviation: 0.01
Normalized holdup BI: 0.63
Normalized holdup BI standard deviation: 0.02
Immobilized partner concentration in holdup experiment (10-6M): 18.55
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: 5.02
Experiment series 4
Protein A protein: DLG1
Protein A fragment: 221-314
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: HPV35-E6
Protein B fragment: 140-149
Protein B site: PBM
Protein B sequence: SKPTRRETEV
Protein B construct: biotin-ttds-SKPTRRETEV
Average holdup BI: 0.47
Normalized holdup BI: 0.61
Immobilized partner concentration in holdup experiment (10-6M): 18.55
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: 4.98
Experiment series 5
Protein A protein: DLG1
Protein A fragment: 221-314
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: HPV35-E6
Protein B fragment: 140-149
Protein B site: PBM
Protein B sequence: SKPTRRETEV
Protein B construct: biotin-ttds-SKPTRRETEV
Average holdup BI: 0.58
Normalized holdup BI: 0.68
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 #6
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 5.15