Details of experiments between Q5TCQ9 and Q13574
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: MAGI3
Protein A fragment: 398-503
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: DGKZ
Protein B fragment: 919-928
Protein B site: PBM
Protein B sequence: IQREDQETAV
Protein B construct: biotin-ttds-IQREDQETAV
Average holdup BI: 0.37
Immobilized partner concentration in holdup experiment (10-6M): 6
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: 5.11
Experiment series 2
Protein A protein: MAGI3
Protein A fragment: 847-941
Protein A site: PDZ_5
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: DGKZ
Protein B fragment: 919-928
Protein B site: PBM
Protein B sequence: IQREDQETAV
Protein B construct: biotin-ttds-IQREDQETAV
Average holdup BI: 0.15
Immobilized partner concentration in holdup experiment (10-6M): 6
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: 4.51
Experiment series 3
Protein A protein: MAGI3
Protein A fragment: 1-114
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: DGKZ
Protein B fragment: 919-928
Protein B site: PBM
Protein B sequence: IQREDQETAV
Protein B construct: biotin-ttds-IQREDQETAV
Average holdup BI: -0.06
Immobilized partner concentration in holdup experiment (10-6M): 6
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: MAGI3
Protein A fragment: 572-655
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: DGKZ
Protein B fragment: 919-928
Protein B site: PBM
Protein B sequence: IQREDQETAV
Protein B construct: biotin-ttds-IQREDQETAV
Average holdup BI: 0.09
Immobilized partner concentration in holdup experiment (10-6M): 6
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: MAGI3
Protein A fragment: 716-815
Protein A site: PDZ_4
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: DGKZ
Protein B fragment: 919-928
Protein B site: PBM
Protein B sequence: IQREDQETAV
Protein B construct: biotin-ttds-IQREDQETAV
Average holdup BI: -0.04
Immobilized partner concentration in holdup experiment (10-6M): 6
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: MAGI3
Protein A fragment: 1014-1103
Protein A site: PDZ_6
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: DGKZ
Protein B fragment: 919-928
Protein B site: PBM
Protein B sequence: IQREDQETAV
Protein B construct: biotin-ttds-IQREDQETAV
Average holdup BI: -0.07
Immobilized partner concentration in holdup experiment (10-6M): 6
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 7
Protein A protein: MAGI3
Protein A fragment: 398-503
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: DGKZ
Protein B fragment: 919-928
Protein B site: PBM
Protein B sequence: IQREDQETAV
Protein B construct: biotin-ttds-IQREDQETAV
Average holdup BI: 0.41
Holdup BI standard deviation: 0.04
Normalized holdup BI: 0.45
Normalized holdup BI standard deviation: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 13.5
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: 2
Measured pKd: 4.85
Experiment series 8
Protein A protein: MAGI3
Protein A fragment: 398-503
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: DGKZ
Protein B fragment: 919-928
Protein B site: PBM
Protein B sequence: IQREDQETAV
Protein B construct: biotin-ttds-IQREDQETAV
Average holdup BI: 0.43
Normalized holdup BI: 0.45
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: 4.71