Details of experiments between Q14160 and K9N5R3
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: SCRIB
Protein A fragment: 1090-1197
Protein A site: PDZ_4
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: MERS-E
Protein B fragment: 71-82
Protein B site: PBM
Protein B sequence: KPPLPPDEWV
Protein B construct: biotin-DSKPPLPPDEWV
Average holdup BI: 0.14
Holdup BI standard deviation: 0.09
Immobilized partner concentration in holdup experiment (10-6M): 20
Experiment method: CALIP_holdup
Details of affinity fitting: BSA and lysozyme internal standards + bacterial cell lysate + normal layout
PUBMED ID of publication: 36115835
Number of measurements: 2
Measured pKd: 3.92
Experiment series 2
Protein A protein: SCRIB
Protein A fragment: 718-814
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: MERS-E
Protein B fragment: 71-82
Protein B site: PBM
Protein B sequence: KPPLPPDEWV
Protein B construct: biotin-DSKPPLPPDEWV
Average holdup BI: -0.05
Holdup BI standard deviation: 0.02
Immobilized partner concentration in holdup experiment (10-6M): 20
Experiment method: CALIP_holdup
Details of affinity fitting: BSA and lysozyme internal standards + bacterial cell lysate + normal layout
PUBMED ID of publication: 36115835
Number of measurements: 2
The affinity was below the detection threshold of the assay.
Experiment series 3
Protein A protein: SCRIB
Protein A fragment: 858-949
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: MERS-E
Protein B fragment: 71-82
Protein B site: PBM
Protein B sequence: KPPLPPDEWV
Protein B construct: biotin-DSKPPLPPDEWV
Average holdup BI: -0.03
Immobilized partner concentration in holdup experiment (10-6M): 20
Experiment method: CALIP_holdup
Details of affinity fitting: BSA and lysozyme internal standards + bacterial cell lysate + 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: SCRIB
Protein A fragment: 999-1093
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: MERS-E
Protein B fragment: 71-82
Protein B site: PBM
Protein B sequence: KPPLPPDEWV
Protein B construct: biotin-DSKPPLPPDEWV
Average holdup BI: -0.06
Holdup BI standard deviation: 0.13
Immobilized partner concentration in holdup experiment (10-6M): 20
Experiment method: CALIP_holdup
Details of affinity fitting: BSA and lysozyme internal standards + bacterial cell lysate + normal layout
PUBMED ID of publication: 36115835
Number of measurements: 2
The affinity was below the detection threshold of the assay.
Experiment series 5
Protein A protein: SCRIB
Protein A fragment: 718-814
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: MERS-E
Protein B fragment: 71-82
Protein B site: PBM
Protein B sequence: KPPLPPDEWV
Protein B construct: biotin-DSKPPLPPDEWV
Average holdup BI: 0.34
Normalized holdup BI: 0.36
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: 4.52
Experiment series 6
Protein A protein: SCRIB
Protein A fragment: 999-1093
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: MERS-E
Protein B fragment: 71-82
Protein B site: PBM
Protein B sequence: KPPLPPDEWV
Protein B construct: biotin-DSKPPLPPDEWV
Average holdup BI: 0.11
Normalized holdup BI: 0.12
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: 3.9
Experiment series 7
Protein A protein: SCRIB
Protein A fragment: 858-949
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: MERS-E
Protein B fragment: 71-82
Protein B site: PBM
Protein B sequence: KPPLPPDEWV
Protein B construct: biotin-DSKPPLPPDEWV
Average holdup BI: 0.05
Holdup BI standard deviation: 0.02
Normalized holdup BI: 0.06
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: 3.58
Experiment series 8
Protein A protein: SCRIB
Protein A fragment: 999-1093
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: MERS-E
Protein B fragment: 73-82
Protein B site: PBM
Protein B sequence: KPPLPPDEWV
Protein B construct: Biotin-ado-ado-KPPLPPDEWV
Average holdup BI: 0.05
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: 3.53
Experiment series 9
Protein A protein: SCRIB
Protein A fragment: 718-814
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: MERS-E
Protein B fragment: 73-82
Protein B site: PBM
Protein B sequence: KPPLPPDEWV
Protein B construct: Biotin-ado-ado-KPPLPPDEWV
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 10
Protein A protein: SCRIB
Protein A fragment: 858-949
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: MERS-E
Protein B fragment: 73-82
Protein B site: PBM
Protein B sequence: KPPLPPDEWV
Protein B construct: Biotin-ado-ado-KPPLPPDEWV
Average holdup BI: 0.03
Normalized holdup BI: 0.03
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.