Details of experiments between Q12959 and Q9UHC3
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: 457-570
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ

Protein B protein: ASIC3
Protein B fragment: 522-531
Protein B site: PBM
Protein B sequence: HRTCYLVTQL
Protein B construct: biotin-ttds-HRTCYLVTQL

Average holdup BI: 0.26
Immobilized partner concentration in holdup experiment (10-6M): 11.5
Experiment method: SAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normal layout
Experimental details: The peptide stock solution had solubility issues. 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: 4.52


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: ASIC3
Protein B fragment: 522-531
Protein B site: PBM
Protein B sequence: HRTCYLVTQL
Protein B construct: biotin-ttds-HRTCYLVTQL

Average holdup BI: 0.12
Immobilized partner concentration in holdup experiment (10-6M): 11.5
Experiment method: SAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normal layout
Experimental details: The peptide stock solution had solubility issues. 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: 4.09


Experiment series 3

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

Protein B protein: ASIC3
Protein B fragment: 522-531
Protein B site: PBM
Protein B sequence: HRTCYLVTQL
Protein B construct: biotin-ttds-HRTCYLVTQL

Average holdup BI: 0.09
Immobilized partner concentration in holdup experiment (10-6M): 11.5
Experiment method: SAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normal layout
Experimental details: The peptide stock solution had solubility issues. 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
The affinity was below the detection threshold of the assay.


Experiment series 4

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

Protein B protein: ASIC3
Protein B fragment: 522-531
Protein B site: PBM
Protein B sequence: HRTCYLVTQL
Protein B construct: biotin-ttds-HRTCYLVTQL

Average holdup BI: 0.44
Normalized holdup BI: 0.53
Immobilized partner concentration in holdup experiment (10-6M): 33
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
Experimental details: The peptide stock solution had solubility issues. 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: 4.56


Experiment series 5

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

Protein B protein: ASIC3
Protein B fragment: 522-531
Protein B site: PBM
Protein B sequence: HRTCYLVTQL
Protein B construct: biotin-ttds-HRTCYLVTQL

Average holdup BI: 0.26
Normalized holdup BI: 0.3
Immobilized partner concentration in holdup experiment (10-6M): 33
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
Experimental details: The peptide stock solution had solubility issues. 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: 4.12


Experiment series 6

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

Protein B protein: ASIC3
Protein B fragment: 522-531
Protein B site: PBM
Protein B sequence: HRTCYLVTQL
Protein B construct: biotin-ttds-HRTCYLVTQL

Average holdup BI: 0.2
Normalized holdup BI: 0.23
Immobilized partner concentration in holdup experiment (10-6M): 33
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
Experimental details: The peptide stock solution had solubility issues. 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: 3.97


Experiment series 7

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

Protein B protein: ASIC3
Protein B fragment: 522-531
Protein B site: PBM
Protein B sequence: HRTCYLVTQL
Protein B construct: Biotin-ado-ado-HRTCYLVTQL

Average holdup BI: 0.19
Normalized holdup BI: 0.23
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: 4.24


Experiment series 8

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

Protein B protein: ASIC3
Protein B fragment: 522-531
Protein B site: PBM
Protein B sequence: HRTCYLVTQL
Protein B construct: Biotin-ado-ado-HRTCYLVTQL

Average holdup BI: 0.24
Normalized holdup BI: 0.27
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: 4.34


Experiment series 9

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

Protein B protein: ASIC3
Protein B fragment: 522-531
Protein B site: PBM
Protein B sequence: HRTCYLVTQL
Protein B construct: Biotin-ado-ado-HRTCYLVTQL

Average holdup BI: 0.39
Normalized holdup BI: 0.58
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: 4.95