Details of experiments between P78352 and Q9NS75
Note that the results of all experiments are listed, regardless of the modification states of the fragments.



Experiment series 1

Protein A protein: DLG4
Protein A fragment: 306-403
Protein A site: PDZ_3
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

Measured dissociation constant (10-6M): 2.51
Standard deviation of dissociation constant (10-6M): 0.13
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit

PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 5.6


Experiment series 2

Protein A protein: DLG4
Protein A fragment: 155-249
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: 0.64
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
Measured pKd: 5.64


Experiment series 3

Protein A protein: DLG4
Protein A fragment: 306-403
Protein A site: PDZ_3
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: 0.59
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
Measured pKd: 5.52


Experiment series 4

Protein A protein: DLG4
Protein A fragment: 62-152
Protein A site: PDZ_1
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: 0.23
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
Measured pKd: 4.71


Experiment series 5

Protein A protein: DLG4
Protein A fragment: 306-403
Protein A site: PDZ_3
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: 0.86
Holdup BI standard deviation: 0.04
Normalized holdup BI: 0.99
Normalized holdup BI standard deviation: 0.05
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: 3
Measured pKd: 6.55


Experiment series 6

Protein A protein: DLG4
Protein A fragment: 155-249
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: 0.66
Holdup BI standard deviation: 0
Normalized holdup BI: 0.85
Normalized holdup BI standard deviation: 0
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: 2
Measured pKd: 5.38


Experiment series 7

Protein A protein: DLG4
Protein A fragment: 62-152
Protein A site: PDZ_1
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: 0.56
Holdup BI standard deviation: 0.05
Normalized holdup BI: 0.67
Normalized holdup BI standard deviation: 0.06
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: 2
Measured pKd: 4.92


Experiment series 8

Protein A protein: DLG4
Protein A fragment: 306-403
Protein A site: PDZ_3
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: 0.8
Normalized holdup BI: 0.94
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: 6.08


Experiment series 9

Protein A protein: DLG4
Protein A fragment: 155-249
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: 0.73
Normalized holdup BI: 0.91
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: 5.83


Experiment series 10

Protein A protein: DLG4
Protein A fragment: 62-152
Protein A site: PDZ_1
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: 0.65
Normalized holdup BI: 0.77
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: 5.34