Details of experiments between Q14CM0 and Q8N448
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: LNX2
Protein B fragment: 334-426
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.1
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: 4.25
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: LNX2
Protein B fragment: 459-560
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.07
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 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: LNX2
Protein B fragment: 592-688
Protein B site: PDZ_4
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.03
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: LNX2
Protein B fragment: 334-426
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.34
Normalized holdup BI: 0.37
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
Measured pKd: 4.47
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: LNX2
Protein B fragment: 459-560
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.01
Normalized holdup BI: 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: 1
The affinity was below the detection threshold of the assay.
Experiment series 6
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: LNX2
Protein B fragment: 334-426
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.54
Normalized holdup BI: 0.54
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: 4.87
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: LNX2
Protein B fragment: 459-560
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.04
Normalized holdup BI: 0.04
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 8
Protein A protein: FRMPD4
Protein A fragment: 67-160
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: LNX2
Protein B fragment: 681-690
Protein B site: PBM
Protein B sequence: TVICWPGSLV
Protein B construct: Biotin-Ahx-TVICWPGSLV
Average holdup BI: 0.01
Holdup BI standard deviation: 0.01
Normalized holdup BI: 0.01
Normalized holdup BI standard deviation: 0.01
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: The peptide stock solution had solubility issues. 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: 2
The affinity was below the detection threshold of the assay.