Details of experiments between Q12959 and P34998
Note that the results of all experiments are listed, regardless of the modification states of the fragments.



Experiment series 1

Protein A protein: DLG1
Protein A fragment: 314-412
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ

Protein B protein: CRHR1
Protein B fragment: 435-444
Protein B site: PBM
Protein B sequence: FHSIKQSTAV
Protein B construct: biotin-ado-ado-FHSIKQSTAV

Average holdup BI: 0.54
Normalized holdup BI: 0.6
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 causing a systematic error in reported affinities.
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 4.98


Experiment series 2

Protein A protein: DLG1
Protein A fragment: 221-314
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ

Protein B protein: CRHR1
Protein B fragment: 435-444
Protein B site: PBM
Protein B sequence: FHSIKQSTAV
Protein B construct: biotin-ado-ado-FHSIKQSTAV

Average holdup BI: 0.25
Normalized holdup BI: 0.29
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 causing a systematic error in reported affinities.
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 4.39


Experiment series 3

Protein A protein: DLG1
Protein A fragment: 457-570
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ

Protein B protein: CRHR1
Protein B fragment: 435-444
Protein B site: PBM
Protein B sequence: FHSIKQSTAV
Protein B construct: biotin-ado-ado-FHSIKQSTAV

Average holdup BI: 0.09
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 causing a systematic error in reported affinities.
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 3.96