Details of experiments between Q96L92 and P06463
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: SNX27
Protein A fragment: 40-141
Protein A site: PDZ
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: 0.28
Holdup BI standard deviation: 0.03
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: 2
Measured pKd: 4.5
Experiment series 2
Protein A protein: SNX27
Protein A fragment: 40-141
Protein A site: PDZ
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
Measured dissociation constant (10-6M): 26.67
Standard deviation of dissociation constant (10-6M): 1.58
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 4.57
Experiment series 3
Protein A protein: SNX27
Protein A fragment: 40-141
Protein A site: PDZ
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: 0.07
Normalized holdup BI: 0.07
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: 3.61
Experiment series 4
Protein A protein: SNX27
Protein A fragment: 40-141
Protein A site: PDZ
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: 0.59
Holdup BI standard deviation: 0.07
Normalized holdup BI: 0.59
Normalized holdup BI standard deviation: 0.07
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: 6
Measured pKd: 4.55
Experiment series 5
Protein A protein: SNX27
Protein A fragment: 40-141
Protein A site: PDZ
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: 0.53
Normalized holdup BI: 0.61
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: 4.59
Experiment series 6
Protein A protein: SNX27
Protein A fragment: 40-141
Protein A site: PDZ
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: 0.53
Normalized holdup BI: 0.56
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: 4.5
Experiment series 7
Protein A protein: SNX27
Protein A fragment: 40-141
Protein A site: PDZ
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: 0.63
Normalized holdup BI: 0.63
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: 5.04