Details of experiments between Q07157 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: TJP1
Protein A fragment: 184-264
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: 0.29
Holdup BI standard deviation: 0.07
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: 4.34
Experiment series 2
Protein A protein: TJP1
Protein A fragment: 14-113
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: 0.01
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: TJP1
Protein A fragment: 415-516
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
Holdup BI standard deviation: 0.06
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 4
Protein A protein: TJP1
Protein A fragment: 184-264
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: 0.17
Normalized holdup BI: 0.2
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: 4.16
Experiment series 5
Protein A protein: TJP1
Protein A fragment: 415-516
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
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
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 3.56
Experiment series 6
Protein A protein: TJP1
Protein A fragment: 184-264
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: 0.17
Normalized holdup BI: 0.21
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: 4.19
Experiment series 7
Protein A protein: TJP1
Protein A fragment: 415-516
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: 0.14
Normalized holdup BI: 0.14
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: 3.96