Details of experiments between Q5T2W1 and Q5SYB0
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: NHERF3
Protein A fragment: 510-519
Protein A site: PBM
Protein A sequence: SSSNSEDTEM
Protein A construct: Biotin-Ahx-SSSNSEDTEM
Protein B protein: FRMPD1
Protein B fragment: 53-142
Protein B site: PDZ
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): 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 2
Protein A protein: NHERF3
Protein A fragment: 2-107
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: FRMPD1
Protein B fragment: 1569-1578
Protein B site: PBM
Protein B sequence: TQKFRASTAL
Protein B construct: Biotin-Ahx-TQKFRASTAL
Average holdup BI: 0.74
Normalized holdup BI: 0.81
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: 5.47
Experiment series 3
Protein A protein: NHERF3
Protein A fragment: 375-459
Protein A site: PDZ_4
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: FRMPD1
Protein B fragment: 1569-1578
Protein B site: PBM
Protein B sequence: TQKFRASTAL
Protein B construct: Biotin-Ahx-TQKFRASTAL
Average holdup BI: 0.36
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: 4.54
Experiment series 4
Protein A protein: NHERF3
Protein A fragment: 238-329
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: FRMPD1
Protein B fragment: 1569-1578
Protein B site: PBM
Protein B sequence: TQKFRASTAL
Protein B construct: Biotin-Ahx-TQKFRASTAL
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
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: NHERF3
Protein A fragment: 2-107
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: FRMPD1
Protein B fragment: 1569-1578
Protein B site: PBM
Protein B sequence: TQKFRASTAL
Protein B construct: Biotin-Ahx-TQKFRASTAL
Average holdup BI: 0.84
Normalized holdup BI: 0.88
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: not reported internal standards of holdup layout #6
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 5.71
Experiment series 6
Protein A protein: NHERF3
Protein A fragment: 375-459
Protein A site: PDZ_4
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: FRMPD1
Protein B fragment: 1569-1578
Protein B site: PBM
Protein B sequence: TQKFRASTAL
Protein B construct: Biotin-Ahx-TQKFRASTAL
Average holdup BI: 0.5
Normalized holdup BI: 0.5
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: not reported internal standards of holdup layout #6
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 4.79
Experiment series 7
Protein A protein: NHERF3
Protein A fragment: 238-329
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: FRMPD1
Protein B fragment: 1569-1578
Protein B site: PBM
Protein B sequence: TQKFRASTAL
Protein B construct: Biotin-Ahx-TQKFRASTAL
Average holdup BI: -0
Normalized holdup BI: -0
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: 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.