Details of experiments between Q86UT5 and Q5KSL6
Note that the results of all experiments are listed, regardless of the modification states of the fragments.



Experiment series 1

Protein A protein: NHERF4
Protein A fragment: 106-208
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ

Protein B protein: DGKK
Protein B fragment: 1262-1271
Protein B site: PBM
Protein B sequence: DPLTPSRSQL
Protein B construct: biotin-ttds-DPLTPSRSQL

Average holdup BI: 0.02
Immobilized partner concentration in holdup experiment (10-6M): 10
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 2

Protein A protein: NHERF4
Protein A fragment: 220-307
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ

Protein B protein: DGKK
Protein B fragment: 1262-1271
Protein B site: PBM
Protein B sequence: DPLTPSRSQL
Protein B construct: biotin-ttds-DPLTPSRSQL

Average holdup BI: -0.04
Immobilized partner concentration in holdup experiment (10-6M): 10
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 3

Protein A protein: NHERF4
Protein A fragment: 325-412
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ

Protein B protein: DGKK
Protein B fragment: 1262-1271
Protein B site: PBM
Protein B sequence: DPLTPSRSQL
Protein B construct: biotin-ttds-DPLTPSRSQL

Average holdup BI: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 10
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 4

Protein A protein: NHERF4
Protein A fragment: 464-550
Protein A site: PDZ_4
Protein A construct: his6-MBP-TEVsite-PDZ

Protein B protein: DGKK
Protein B fragment: 1262-1271
Protein B site: PBM
Protein B sequence: DPLTPSRSQL
Protein B construct: biotin-ttds-DPLTPSRSQL

Average holdup BI: -0.02
Immobilized partner concentration in holdup experiment (10-6M): 10
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: NHERF4
Protein A fragment: 106-208
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ

Protein B protein: DGKK
Protein B fragment: 1262-1271
Protein B site: PBM
Protein B sequence: DPLTPSRSQL
Protein B construct: biotin-ttds-DPLTPSRSQL

Average holdup BI: 0.1
Normalized holdup BI: 0.1
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
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 3.81


Experiment series 6

Protein A protein: NHERF4
Protein A fragment: 220-307
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ

Protein B protein: DGKK
Protein B fragment: 1262-1271
Protein B site: PBM
Protein B sequence: DPLTPSRSQL
Protein B construct: biotin-ttds-DPLTPSRSQL

Average holdup BI: 0.01
Normalized holdup BI: 0.01
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
PUBMED ID of publication: 36115835
Number of measurements: 1
The affinity was below the detection threshold of the assay.


Experiment series 7

Protein A protein: NHERF4
Protein A fragment: 464-550
Protein A site: PDZ_4
Protein A construct: his6-MBP-TEVsite-PDZ

Protein B protein: DGKK
Protein B fragment: 1262-1271
Protein B site: PBM
Protein B sequence: DPLTPSRSQL
Protein B construct: biotin-ttds-DPLTPSRSQL

Average holdup BI: 0.01
Normalized holdup BI: 0.01
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
PUBMED ID of publication: 36115835
Number of measurements: 1
The affinity was below the detection threshold of the assay.