Details of experiments between the fragments O15018~326-424 and Q9NS75~337-346
Note that the results of all experiments are listed, regardless of the modification states of the fragments.



Experiment series 1

Protein A protein: PDZD2
Protein A fragment: 326-424
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ

Protein B protein: CYSLTR2
Protein B fragment: 337-346
Protein B site: PBM
Protein B sequence: SVWLRKETRV
Protein B construct: biotin-ttds-SVWLRKETRV

Average holdup BI: -0.11
Immobilized partner concentration in holdup experiment (10-6M): 6.7
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 2

Protein A protein: PDZD2
Protein A fragment: 326-424
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ

Protein B protein: CYSLTR2
Protein B fragment: 337-346
Protein B site: PBM
Protein B sequence: SVWLRKETRV
Protein B construct: biotin-ttds-SVWLRKETRV

Average holdup BI: 0.14
Normalized holdup BI: 0.14
Immobilized partner concentration in holdup experiment (10-6M): 27
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: PDZD2
Protein A fragment: 326-424
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ

Protein B protein: CYSLTR2
Protein B fragment: 337-346
Protein B site: PBM
Protein B sequence: SVWLRKETRV
Protein B construct: Biotin-ado-ado-SVWLRKETRV

Average holdup BI: 0.17
Normalized holdup BI: 0.17
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: 4.07