Details of experiments between Q13574 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: DGKZ
Protein A fragment: 919-928
Protein A site: PBM
Protein A sequence: IQREDQETAV
Protein A construct: biotin-ttds-IQREDQETAV

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): 151.39
Standard deviation of dissociation constant (10-6M): 21.64
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit

PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 3.82


Experiment series 2

Protein A protein: DGKZ
Protein A fragment: 919-928
Protein A site: PBM
Protein A sequence: IQREDQETAV
Protein A construct: biotin-ttds-IQREDQETAV

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

Average holdup BI: 0.25
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: 4.83


Experiment series 3

Protein A protein: DGKZ
Protein A fragment: 919-928
Protein A site: PBM
Protein A sequence: IQREDQETAV
Protein A construct: biotin-ttds-IQREDQETAV

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

Average holdup BI: 0.11
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: 4.34


Experiment series 4

Protein A protein: DGKZ
Protein A fragment: 919-928
Protein A site: PBM
Protein A sequence: IQREDQETAV
Protein A construct: biotin-ttds-IQREDQETAV

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

Average holdup BI: 0.08
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: DGKZ
Protein A fragment: 919-928
Protein A site: PBM
Protein A sequence: IQREDQETAV
Protein A construct: biotin-ttds-IQREDQETAV

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

Average holdup BI: 0.39
Holdup BI standard deviation: 0.01
Normalized holdup BI: 0.51
Normalized holdup BI standard deviation: 0.02
Immobilized partner concentration in holdup experiment (10-6M): 13.5
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: 4.96


Experiment series 6

Protein A protein: DGKZ
Protein A fragment: 919-928
Protein A site: PBM
Protein A sequence: IQREDQETAV
Protein A construct: biotin-ttds-IQREDQETAV

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

Average holdup BI: 0.32
Holdup BI standard deviation: 0
Normalized holdup BI: 0.39
Normalized holdup BI standard deviation: 0
Immobilized partner concentration in holdup experiment (10-6M): 13.5
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: 4.72


Experiment series 7

Protein A protein: DGKZ
Protein A fragment: 919-928
Protein A site: PBM
Protein A sequence: IQREDQETAV
Protein A construct: biotin-ttds-IQREDQETAV

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

Average holdup BI: 0.07
Holdup BI standard deviation: 0.03
Normalized holdup BI: 0.08
Normalized holdup BI standard deviation: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 13.5
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: 4
Measured pKd: 3.84


Experiment series 8

Protein A protein: DGKZ
Protein A fragment: 919-928
Protein A site: PBM
Protein A sequence: IQREDQETAV
Protein A construct: biotin-ttds-IQREDQETAV

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
Normalized holdup BI: 0.07
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: not reported internal standards of holdup layout #6
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 3.59


Experiment series 9

Protein A protein: DGKZ
Protein A fragment: 919-928
Protein A site: PBM
Protein A sequence: IQREDQETAV
Protein A construct: biotin-ttds-IQREDQETAV

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

Average holdup BI: 0.39
Normalized holdup BI: 0.48
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: not reported internal standards of holdup layout #6
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 4.76


Experiment series 10

Protein A protein: DGKZ
Protein A fragment: 919-928
Protein A site: PBM
Protein A sequence: IQREDQETAV
Protein A construct: biotin-ttds-IQREDQETAV

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

Average holdup BI: 0.29
Normalized holdup BI: 0.34
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: not reported internal standards of holdup layout #6
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 4.5