Details of experiments between Q5T2W1 and Q9Y6N9
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: 2-107
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ

Protein B protein: USH1C
Protein B fragment: 543-552
Protein B site: PBM
Protein B sequence: KEYDDELTFF
Protein B construct: Biotin-Ahx-KEYDDELTFF

Average holdup BI: 0.22
Normalized holdup BI: 0.24
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: 4.27


Experiment series 2

Protein A protein: NHERF3
Protein A fragment: 375-459
Protein A site: PDZ_4
Protein A construct: his6-MBP-TEVsite-PDZ

Protein B protein: USH1C
Protein B fragment: 543-552
Protein B site: PBM
Protein B sequence: KEYDDELTFF
Protein B construct: Biotin-Ahx-KEYDDELTFF

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
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 3.8


Experiment series 3

Protein A protein: NHERF3
Protein A fragment: 238-329
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ

Protein B protein: USH1C
Protein B fragment: 543-552
Protein B site: PBM
Protein B sequence: KEYDDELTFF
Protein B construct: Biotin-Ahx-KEYDDELTFF

Average holdup BI: 0.02
Normalized holdup BI: 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: 1
The affinity was below the detection threshold of the assay.