Details of experiments between Q14160 and P26554
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: SCRIB
Protein A fragment: 718-814
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: HPV51-E6
Protein B fragment: 142-151
Protein B site: PBM
Protein B sequence: RTRQRNETQV
Protein B construct: biotin-ttds-RTRQRNETQV
Measured dissociation constant (10-6M): 30.64
Standard deviation of dissociation constant (10-6M): 0.88
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 4.51
Experiment series 2
Protein A protein: SCRIB
Protein A fragment: 999-1093
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: HPV51-E6
Protein B fragment: 142-151
Protein B site: PBM
Protein B sequence: RTRQRNETQV
Protein B construct: biotin-ttds-RTRQRNETQV
Average holdup BI: 0.35
Immobilized partner concentration in holdup experiment (10-6M): 7.7
Experiment method: SAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normal layout
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 4.93
Experiment series 3
Protein A protein: SCRIB
Protein A fragment: 718-814
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: HPV51-E6
Protein B fragment: 142-151
Protein B site: PBM
Protein B sequence: RTRQRNETQV
Protein B construct: biotin-ttds-RTRQRNETQV
Average holdup BI: 0.26
Immobilized partner concentration in holdup experiment (10-6M): 7.7
Experiment method: SAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normal layout
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 4.72
Experiment series 4
Protein A protein: SCRIB
Protein A fragment: 1090-1197
Protein A site: PDZ_4
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: HPV51-E6
Protein B fragment: 142-151
Protein B site: PBM
Protein B sequence: RTRQRNETQV
Protein B construct: biotin-ttds-RTRQRNETQV
Average holdup BI: 0.12
Immobilized partner concentration in holdup experiment (10-6M): 7.7
Experiment method: SAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normal layout
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 4.27
Experiment series 5
Protein A protein: SCRIB
Protein A fragment: 858-949
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: HPV51-E6
Protein B fragment: 142-151
Protein B site: PBM
Protein B sequence: RTRQRNETQV
Protein B construct: biotin-ttds-RTRQRNETQV
Average holdup BI: 0.01
Immobilized partner concentration in holdup experiment (10-6M): 7.7
Experiment method: SAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normal 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: SCRIB
Protein A fragment: 718-814
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: HPV51-E6
Protein B fragment: 142-151
Protein B site: PBM
Protein B sequence: RTRQRNETQV
Protein B construct: biotin-ttds-RTRQRNETQV
Average holdup BI: 0.4
Holdup BI standard deviation: 0.04
Normalized holdup BI: 0.42
Normalized holdup BI standard deviation: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 21.5
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: 4
Measured pKd: 4.56
Experiment series 7
Protein A protein: SCRIB
Protein A fragment: 858-949
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: HPV51-E6
Protein B fragment: 142-151
Protein B site: PBM
Protein B sequence: RTRQRNETQV
Protein B construct: biotin-ttds-RTRQRNETQV
Average holdup BI: 0.27
Normalized holdup BI: 0.33
Immobilized partner concentration in holdup experiment (10-6M): 21.5
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: 4.39
Experiment series 8
Protein A protein: SCRIB
Protein A fragment: 999-1093
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: HPV51-E6
Protein B fragment: 142-151
Protein B site: PBM
Protein B sequence: RTRQRNETQV
Protein B construct: biotin-ttds-RTRQRNETQV
Average holdup BI: 0.21
Normalized holdup BI: 0.23
Immobilized partner concentration in holdup experiment (10-6M): 21.5
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: 4.17