Details of experiments between Q96QZ7 and Q14CM0
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: MAGI1
Protein A fragment: 638-725
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: FRMPD4
Protein B fragment: 1313-1322
Protein B site: PBM
Protein B sequence: RLPKIKETTV
Protein B construct: biotin-ttds-RLPKIKETTV
Average holdup BI: 0.27
Immobilized partner concentration in holdup experiment (10-6M): 6.6
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: 4.82
Experiment series 2
Protein A protein: MAGI1
Protein A fragment: 1146-1238
Protein A site: PDZ_6
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: FRMPD4
Protein B fragment: 1313-1322
Protein B site: PBM
Protein B sequence: RLPKIKETTV
Protein B construct: biotin-ttds-RLPKIKETTV
Average holdup BI: 0.18
Immobilized partner concentration in holdup experiment (10-6M): 6.6
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: 4.57
Experiment series 3
Protein A protein: MAGI1
Protein A fragment: 986-1098
Protein A site: PDZ_5
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: FRMPD4
Protein B fragment: 1313-1322
Protein B site: PBM
Protein B sequence: RLPKIKETTV
Protein B construct: biotin-ttds-RLPKIKETTV
Average holdup BI: 0.16
Immobilized partner concentration in holdup experiment (10-6M): 6.6
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: 4.5
Experiment series 4
Protein A protein: MAGI1
Protein A fragment: 456-580
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: FRMPD4
Protein B fragment: 1313-1322
Protein B site: PBM
Protein B sequence: RLPKIKETTV
Protein B construct: biotin-ttds-RLPKIKETTV
Average holdup BI: 0.14
Immobilized partner concentration in holdup experiment (10-6M): 6.6
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: 4.43
Experiment series 5
Protein A protein: MAGI1
Protein A fragment: 1-106
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: FRMPD4
Protein B fragment: 1313-1322
Protein B site: PBM
Protein B sequence: RLPKIKETTV
Protein B construct: biotin-ttds-RLPKIKETTV
Average holdup BI: 0.06
Immobilized partner concentration in holdup experiment (10-6M): 6.6
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: MAGI1
Protein A fragment: 836-926
Protein A site: PDZ_4
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: FRMPD4
Protein B fragment: 1313-1322
Protein B site: PBM
Protein B sequence: RLPKIKETTV
Protein B construct: biotin-ttds-RLPKIKETTV
Average holdup BI: 0.04
Immobilized partner concentration in holdup experiment (10-6M): 6.6
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: MAGI1
Protein A fragment: 456-580
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: FRMPD4
Protein B fragment: 1313-1322
Protein B site: PBM
Protein B sequence: RLPKIKETTV
Protein B construct: biotin-ttds-RLPKIKETTV
Average holdup BI: 0.34
Holdup BI standard deviation: 0.02
Normalized holdup BI: 0.39
Normalized holdup BI standard deviation: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 21.6
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: 4.49
Experiment series 8
Protein A protein: MAGI1
Protein A fragment: 1-106
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: FRMPD4
Protein B fragment: 1313-1322
Protein B site: PBM
Protein B sequence: RLPKIKETTV
Protein B construct: biotin-ttds-RLPKIKETTV
Average holdup BI: -0
Normalized holdup BI: -0
Immobilized partner concentration in holdup experiment (10-6M): 21.6
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
The affinity was below the detection threshold of the assay.
Experiment series 9
Protein A protein: MAGI1
Protein A fragment: 1-106
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: FRMPD4
Protein B fragment: 1313-1322
Protein B site: PBM
Protein B sequence: RLPKIKETTV
Protein B construct: Biotin-ado-ado-RLPKIKETTV
Average holdup BI: 0.08
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: 3.71
Experiment series 10
Protein A protein: MAGI1
Protein A fragment: 456-580
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: FRMPD4
Protein B fragment: 1313-1322
Protein B site: PBM
Protein B sequence: RLPKIKETTV
Protein B construct: Biotin-ado-ado-RLPKIKETTV
Average holdup BI: 0.43
Normalized holdup BI: 0.48
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: 4.76