Details of experiments between O75084 and P78352
Note that the results of all experiments are listed, regardless of the modification states of the fragments.



Experiment series 1

Protein A protein: FZD7
Protein A fragment: 565-574
Protein A site: PBM
Protein A sequence: SHSSKGETAV
Protein A construct: biotin-ttds-SHSSKGETAV

Protein B protein: DLG4
Protein B fragment: 306-403
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ

Measured dissociation constant (10-6M): 82.13
Standard deviation of dissociation constant (10-6M): 10.67
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit

PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 4.09


Experiment series 2

Protein A protein: FZD7
Protein A fragment: 565-574
Protein A site: PBM
Protein A sequence: SHSSKGETAV
Protein A construct: biotin-ttds-SHSSKGETAV

Protein B protein: DLG4
Protein B fragment: 62-152
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.13
Immobilized partner concentration in holdup experiment (10-6M): 6
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: 4.43


Experiment series 3

Protein A protein: FZD7
Protein A fragment: 565-574
Protein A site: PBM
Protein A sequence: SHSSKGETAV
Protein A construct: biotin-ttds-SHSSKGETAV

Protein B protein: DLG4
Protein B fragment: 155-249
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.1
Immobilized partner concentration in holdup experiment (10-6M): 6
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: 4.32


Experiment series 4

Protein A protein: FZD7
Protein A fragment: 565-574
Protein A site: PBM
Protein A sequence: SHSSKGETAV
Protein A construct: biotin-ttds-SHSSKGETAV

Protein B protein: DLG4
Protein B fragment: 306-403
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.06
Immobilized partner concentration in holdup experiment (10-6M): 6
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 5

Protein A protein: FZD7
Protein A fragment: 565-574
Protein A site: PBM
Protein A sequence: SHSSKGETAV
Protein A construct: biotin-ttds-SHSSKGETAV

Protein B protein: DLG4
Protein B fragment: 155-249
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.38
Normalized holdup BI: 0.49
Immobilized partner concentration in holdup experiment (10-6M): 21.5
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: 4.69


Experiment series 6

Protein A protein: FZD7
Protein A fragment: 565-574
Protein A site: PBM
Protein A sequence: SHSSKGETAV
Protein A construct: biotin-ttds-SHSSKGETAV

Protein B protein: DLG4
Protein B fragment: 62-152
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.28
Normalized holdup BI: 0.33
Immobilized partner concentration in holdup experiment (10-6M): 21.5
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: 4.4


Experiment series 7

Protein A protein: FZD7
Protein A fragment: 565-574
Protein A site: PBM
Protein A sequence: SHSSKGETAV
Protein A construct: biotin-ttds-SHSSKGETAV

Protein B protein: DLG4
Protein B fragment: 306-403
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.16
Holdup BI standard deviation: 0.09
Normalized holdup BI: 0.18
Normalized holdup BI standard deviation: 0.11
Immobilized partner concentration in holdup experiment (10-6M): 21.5
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: 4.02