Details of experiments between Q86UL8 and P20020
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: MAGI2
Protein A fragment: 915-1017
Protein A site: PDZ_5
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: ATP2B1
Protein B fragment: 1211-1220
Protein B site: PBM
Protein B sequence: SPLHSLETSL
Protein B construct: biotin-ttds-SPLHSLETSL
Average holdup BI: 0.26
Holdup BI standard deviation: 0.02
Normalized holdup BI: 0.26
Normalized holdup BI standard deviation: 0.02
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: 2
Measured pKd: 4.31
Experiment series 2
Protein A protein: MAGI2
Protein A fragment: 1141-1229
Protein A site: PDZ_6
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: ATP2B1
Protein B fragment: 1211-1220
Protein B site: PBM
Protein B sequence: SPLHSLETSL
Protein B construct: biotin-ttds-SPLHSLETSL
Average holdup BI: 0.19
Holdup BI standard deviation: 0
Normalized holdup BI: 0.23
Normalized holdup BI standard deviation: 0
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: 2
Measured pKd: 4.24
Experiment series 3
Protein A protein: MAGI2
Protein A fragment: 410-522
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: ATP2B1
Protein B fragment: 1211-1220
Protein B site: PBM
Protein B sequence: SPLHSLETSL
Protein B construct: biotin-ttds-SPLHSLETSL
Average holdup BI: 0.1
Normalized holdup BI: 0.11
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: 3.86
Experiment series 4
Protein A protein: MAGI2
Protein A fragment: 600-682
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: ATP2B1
Protein B fragment: 1211-1220
Protein B site: PBM
Protein B sequence: SPLHSLETSL
Protein B construct: biotin-ttds-SPLHSLETSL
Average holdup BI: 0.04
Holdup BI standard deviation: 0.06
Normalized holdup BI: 0.05
Normalized holdup BI standard deviation: 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: 2
Measured pKd: 3.5
Experiment series 5
Protein A protein: MAGI2
Protein A fragment: 772-865
Protein A site: PDZ_4
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: ATP2B1
Protein B fragment: 1211-1220
Protein B site: PBM
Protein B sequence: SPLHSLETSL
Protein B construct: biotin-ttds-SPLHSLETSL
Average holdup BI: 0.04
Holdup BI standard deviation: 0.02
Normalized holdup BI: 0.05
Normalized holdup BI standard deviation: 0.02
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: 2
Measured pKd: 3.48
Experiment series 6
Protein A protein: MAGI2
Protein A fragment: 410-522
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: ATP2B1
Protein B fragment: 1211-1220
Protein B site: PBM
Protein B sequence: SPLHSLETSL
Protein B construct: Biotin-ado-ado-SPLHSLETSL
Average holdup BI: 0.07
Normalized holdup BI: 0.08
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: Crude peptide synthesis product. The peptide concentration in the affinity conversion step may differ from reality. Not reported internal standards of holdup layout #6
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 3.7
Experiment series 7
Protein A protein: MAGI2
Protein A fragment: 1141-1229
Protein A site: PDZ_6
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: ATP2B1
Protein B fragment: 1211-1220
Protein B site: PBM
Protein B sequence: SPLHSLETSL
Protein B construct: Biotin-ado-ado-SPLHSLETSL
Average holdup BI: 0.29
Normalized holdup BI: 0.29
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: Crude peptide synthesis product. The peptide concentration in the affinity conversion step may differ from reality. Not reported internal standards of holdup layout #6
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 4.39
Experiment series 8
Protein A protein: MAGI2
Protein A fragment: 600-682
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: ATP2B1
Protein B fragment: 1211-1220
Protein B site: PBM
Protein B sequence: SPLHSLETSL
Protein B construct: Biotin-ado-ado-SPLHSLETSL
Average holdup BI: 0.06
Normalized holdup BI: 0.06
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: Crude peptide synthesis product. The peptide concentration in the affinity conversion step may differ from reality. Not reported internal standards of holdup layout #6
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 3.56
Experiment series 9
Protein A protein: MAGI2
Protein A fragment: 772-865
Protein A site: PDZ_4
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: ATP2B1
Protein B fragment: 1211-1220
Protein B site: PBM
Protein B sequence: SPLHSLETSL
Protein B construct: Biotin-ado-ado-SPLHSLETSL
Average holdup BI: 0.04
Normalized holdup BI: 0.06
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: Crude peptide synthesis product. The peptide concentration in the affinity conversion step may differ from reality. Not reported internal standards of holdup layout #6
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 3.52
Experiment series 10
Protein A protein: MAGI2
Protein A fragment: 915-1017
Protein A site: PDZ_5
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: ATP2B1
Protein B fragment: 1211-1220
Protein B site: PBM
Protein B sequence: SPLHSLETSL
Protein B construct: Biotin-ado-ado-SPLHSLETSL
Average holdup BI: 0.31
Normalized holdup BI: 0.31
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: Crude peptide synthesis product. The peptide concentration in the affinity conversion step may differ from reality. Not reported internal standards of holdup layout #6
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 4.44