Details of experiments between Q8TBB1 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: LNX1
Protein A fragment: 719-728
Protein A site: PBM
Protein A sequence: TIVSWPGTFL
Protein A construct: Biotin-Ahx-TIVSWPGTFL
Protein B protein: FRMPD4
Protein B fragment: 67-160
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.08
Holdup BI standard deviation: 0.01
Normalized holdup BI: 0.09
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: 3.73
Experiment series 2
Protein A protein: LNX1
Protein A fragment: 371-470
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: 0.15
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 3
Protein A protein: LNX1
Protein A fragment: 261-371
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: -0.04
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 4
Protein A protein: LNX1
Protein A fragment: 500-596
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
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 5
Protein A protein: LNX1
Protein A fragment: 371-470
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: 0.05
Normalized holdup BI: 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: 1
Measured pKd: 3.42
Experiment series 6
Protein A protein: LNX1
Protein A fragment: 261-371
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: 0.04
Normalized holdup BI: 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: 1
The affinity was below the detection threshold of the assay.
Experiment series 7
Protein A protein: LNX1
Protein A fragment: 500-596
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: 0.01
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 8
Protein A protein: LNX1
Protein A fragment: 371-470
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: 0.1
Normalized holdup BI: 0.1
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: 3.81
Experiment series 9
Protein A protein: LNX1
Protein A fragment: 500-596
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-ado-ado-RLPKIKETTV
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
The affinity was below the detection threshold of the assay.
Experiment series 10
Protein A protein: LNX1
Protein A fragment: 261-371
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: 0.13
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: 3.93