Details of experiments between Q14CM0 and Q9P202
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: FRMPD4
Protein A fragment: 1313-1322
Protein A site: PBM
Protein A sequence: RLPKIKETTV
Protein A construct: biotin-ttds-RLPKIKETTV
Protein B protein: DFNB31
Protein B fragment: 263-379
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.15
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: 4.46
Experiment series 2
Protein A protein: FRMPD4
Protein A fragment: 1313-1322
Protein A site: PBM
Protein A sequence: RLPKIKETTV
Protein A construct: biotin-ttds-RLPKIKETTV
Protein B protein: DFNB31
Protein B fragment: 136-226
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.12
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: 4.36
Experiment series 3
Protein A protein: FRMPD4
Protein A fragment: 1313-1322
Protein A site: PBM
Protein A sequence: RLPKIKETTV
Protein A construct: biotin-ttds-RLPKIKETTV
Protein B protein: DFNB31
Protein B fragment: 813-904
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.01
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 4
Protein A protein: FRMPD4
Protein A fragment: 1313-1322
Protein A site: PBM
Protein A sequence: RLPKIKETTV
Protein A construct: biotin-ttds-RLPKIKETTV
Protein B protein: DFNB31
Protein B fragment: 263-379
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.22
Holdup BI standard deviation: 0.05
Normalized holdup BI: 0.22
Normalized holdup BI standard deviation: 0.05
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: 4.12
Experiment series 5
Protein A protein: FRMPD4
Protein A fragment: 1313-1322
Protein A site: PBM
Protein A sequence: RLPKIKETTV
Protein A construct: biotin-ttds-RLPKIKETTV
Protein B protein: DFNB31
Protein B fragment: 136-226
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.16
Holdup BI standard deviation: 0.04
Normalized holdup BI: 0.16
Normalized holdup BI standard deviation: 0.04
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: 3.96
Experiment series 6
Protein A protein: FRMPD4
Protein A fragment: 1313-1322
Protein A site: PBM
Protein A sequence: RLPKIKETTV
Protein A construct: biotin-ttds-RLPKIKETTV
Protein B protein: DFNB31
Protein B fragment: 813-904
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.03
Holdup BI standard deviation: 0.01
Normalized holdup BI: 0.03
Normalized holdup BI standard deviation: 0.01
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
The affinity was below the detection threshold of the assay.
Experiment series 7
Protein A protein: FRMPD4
Protein A fragment: 1313-1322
Protein A site: PBM
Protein A sequence: RLPKIKETTV
Protein A construct: Biotin-ado-ado-RLPKIKETTV
Protein B protein: DFNB31
Protein B fragment: 263-379
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.43
Normalized holdup BI: 0.43
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: 4.66
Experiment series 8
Protein A protein: FRMPD4
Protein A fragment: 1313-1322
Protein A site: PBM
Protein A sequence: RLPKIKETTV
Protein A construct: Biotin-ado-ado-RLPKIKETTV
Protein B protein: DFNB31
Protein B fragment: 813-904
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: -0.09
Normalized holdup BI: -0.09
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
The affinity was below the detection threshold of the assay.
Experiment series 9
Protein A protein: FRMPD4
Protein A fragment: 1313-1322
Protein A site: PBM
Protein A sequence: RLPKIKETTV
Protein A construct: Biotin-ado-ado-RLPKIKETTV
Protein B protein: DFNB31
Protein B fragment: 136-226
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.33
Normalized holdup BI: 0.33
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: 4.47
Experiment series 10
Protein A protein: FRMPD4
Protein A fragment: 67-160
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: DFNB31
Protein B fragment: 565-578
Protein B site: Internal_PBM
Protein B construct: Biotin-GSGSGS-VSVDDVRSTSQGLSSY-CONH2
Average holdup BI: -0.02
Normalized holdup BI: -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: 1
The affinity was below the detection threshold of the assay.
Experiment series 11
Protein A protein: FRMPD4
Protein A fragment: 67-160
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: DFNB31
Protein B fragment: 898-907
Protein B site: PBM
Protein B sequence: FLVTEFNVML
Protein B construct: biotin-ado-ado-FLVTEFNVML
Average holdup BI: 0.56
Normalized holdup BI: 0.56
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 causing a systematic error in reported affinities.
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 4.9