Details of experiments between Q14160 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: SCRIB
Protein A fragment: 718-814
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

Measured dissociation constant (10-6M): 46.02
Standard deviation of dissociation constant (10-6M): 7.93
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit

PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 4.34


Experiment series 2

Protein A protein: SCRIB
Protein A fragment: 1090-1197
Protein A site: PDZ_4
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: 0.31
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: 4.91


Experiment series 3

Protein A protein: SCRIB
Protein A fragment: 999-1093
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: 0.21
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: 4.66


Experiment series 4

Protein A protein: SCRIB
Protein A fragment: 718-814
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: 0.14
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: 4.41


Experiment series 5

Protein A protein: SCRIB
Protein A fragment: 858-949
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: 0.01
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 6

Protein A protein: SCRIB
Protein A fragment: 999-1093
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: 0.57
Normalized holdup BI: 0.63
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: 4.85


Experiment series 7

Protein A protein: SCRIB
Protein A fragment: 858-949
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: 0.32
Holdup BI standard deviation: 0
Normalized holdup BI: 0.4
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: 4.42


Experiment series 8

Protein A protein: SCRIB
Protein A fragment: 718-814
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: 0.31
Holdup BI standard deviation: 0.07
Normalized holdup BI: 0.33
Normalized holdup BI standard deviation: 0.07
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: 7
Measured pKd: 4.28


Experiment series 9

Protein A protein: SCRIB
Protein A fragment: 999-1093
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: 0.44
Normalized holdup BI: 0.51
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: 4.82


Experiment series 10

Protein A protein: SCRIB
Protein A fragment: 718-814
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: 0.46
Normalized holdup BI: 0.46
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: 4.73


Experiment series 11

Protein A protein: SCRIB
Protein A fragment: 858-949
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: 0.49
Normalized holdup BI: 0.49
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: 4.79