Details of experiments between the fragments Q15700~418-513 and P06463~149-158
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: DLG2
Protein A fragment: 418-513
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: HPV18-E6
Protein B fragment: 149-158
Protein B site: PBM
Protein B sequence: RLQRRRETQV
Protein B construct: biotin-ttds-RLQRRRETQV
Average holdup BI: 0.23
Immobilized partner concentration in holdup experiment (10-6M): 13.3
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: 4.38
Experiment series 2
Protein A protein: DLG2
Protein A fragment: 418-513
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: HPV18-E6
Protein B fragment: 149-158
Protein B site: PBM
Protein B modification: (TPO)156
Protein B sequence: RLQRRRETQV
Protein B construct: Biotin-ttds-RLQRRREpTQV
Average holdup BI: 0.12
Normalized holdup BI: 0.12
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: 3.88
Experiment series 3
Protein A protein: DLG2
Protein A fragment: 418-513
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: HPV18-E6
Protein B fragment: 149-158
Protein B site: PBM
Protein B sequence: RLQRRRETQV
Protein B construct: biotin-ttds-RLQRRRETQV
Average holdup BI: 0.82
Holdup BI standard deviation: 0.02
Normalized holdup BI: 0.82
Normalized holdup BI standard deviation: 0.02
Immobilized partner concentration in holdup experiment (10-6M): 43
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: 5.06
Experiment series 4
Protein A protein: DLG2
Protein A fragment: 418-513
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: HPV18-E6
Protein B fragment: 149-158
Protein B site: PBM
Protein B sequence: RLQRRRETQV
Protein B construct: biotin-ttds-RLQRRRETQV
Average holdup BI: 0.82
Normalized holdup BI: 0.82
Immobilized partner concentration in holdup experiment (10-6M): 43
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: DLG2
Protein A fragment: 418-513
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: HPV18-E6
Protein B fragment: 149-158
Protein B site: PBM
Protein B sequence: RLQRRRETQV
Protein B construct: biotin-ttds-RLQRRRETQV
Average holdup BI: 0.82
Normalized holdup BI: 0.82
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: 5.51