Details of experiments between P78352 and A4U7F2
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: DLG4
Protein A fragment: 306-403
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: HPV67-E6
Protein B fragment: 140-149
Protein B site: PBM
Protein B sequence: SRPQRTQTQV
Protein B construct: biotin-ttds-SRPQRTQTQV
Measured dissociation constant (10-6M): 33.89
Standard deviation of dissociation constant (10-6M): 3.22
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 4.47
Experiment series 2
Protein A protein: DLG4
Protein A fragment: 155-249
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: HPV67-E6
Protein B fragment: 140-149
Protein B site: PBM
Protein B sequence: SRPQRTQTQV
Protein B construct: biotin-ttds-SRPQRTQTQV
Average holdup BI: 0.46
Immobilized partner concentration in holdup experiment (10-6M): 10
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: 5.01
Experiment series 3
Protein A protein: DLG4
Protein A fragment: 62-152
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: HPV67-E6
Protein B fragment: 140-149
Protein B site: PBM
Protein B sequence: SRPQRTQTQV
Protein B construct: biotin-ttds-SRPQRTQTQV
Average holdup BI: 0
Immobilized partner concentration in holdup experiment (10-6M): 10
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 4
Protein A protein: DLG4
Protein A fragment: 306-403
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: HPV67-E6
Protein B fragment: 140-149
Protein B site: PBM
Protein B sequence: SRPQRTQTQV
Protein B construct: biotin-ttds-SRPQRTQTQV
Average holdup BI: -0.04
Immobilized partner concentration in holdup experiment (10-6M): 10
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 5
Protein A protein: DLG4
Protein A fragment: 155-249
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: HPV67-E6
Protein B fragment: 140-149
Protein B site: PBM
Protein B sequence: SRPQRTQTQV
Protein B construct: biotin-ttds-SRPQRTQTQV
Average holdup BI: 0.49
Normalized holdup BI: 0.64
Immobilized partner concentration in holdup experiment (10-6M): 22
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: 4.96
Experiment series 6
Protein A protein: DLG4
Protein A fragment: 62-152
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: HPV67-E6
Protein B fragment: 140-149
Protein B site: PBM
Protein B sequence: SRPQRTQTQV
Protein B construct: biotin-ttds-SRPQRTQTQV
Average holdup BI: 0.36
Normalized holdup BI: 0.43
Immobilized partner concentration in holdup experiment (10-6M): 22
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: 4.58
Experiment series 7
Protein A protein: DLG4
Protein A fragment: 306-403
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: HPV67-E6
Protein B fragment: 140-149
Protein B site: PBM
Protein B sequence: SRPQRTQTQV
Protein B construct: biotin-ttds-SRPQRTQTQV
Average holdup BI: 0.23
Holdup BI standard deviation: 0.03
Normalized holdup BI: 0.26
Normalized holdup BI standard deviation: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 22
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: 4.23