Details of experiments between P78352 and Q15311
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: RALBP1
Protein B fragment: 646-655
Protein B site: PBM
Protein B sequence: PSRDRKETSI
Protein B construct: biotin-ttds-PSRDRKETSI

Measured dissociation constant (10-6M): 10.2
Standard deviation of dissociation constant (10-6M): 0.81
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit

PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 4.99


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: RALBP1
Protein B fragment: 646-655
Protein B site: PBM
Protein B sequence: PSRDRKETSI
Protein B construct: biotin-ttds-PSRDRKETSI

Average holdup BI: 0.41
Immobilized partner concentration in holdup experiment (10-6M): 20
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: 4.59


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: RALBP1
Protein B fragment: 646-655
Protein B site: PBM
Protein B sequence: PSRDRKETSI
Protein B construct: biotin-ttds-PSRDRKETSI

Average holdup BI: 0.27
Immobilized partner concentration in holdup experiment (10-6M): 20
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: 4.3


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: RALBP1
Protein B fragment: 646-655
Protein B site: PBM
Protein B sequence: PSRDRKETSI
Protein B construct: biotin-ttds-PSRDRKETSI

Average holdup BI: 0.09
Immobilized partner concentration in holdup experiment (10-6M): 20
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: DLG4
Protein A fragment: 306-403
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ

Protein B protein: RALBP1
Protein B fragment: 646-655
Protein B site: PBM
Protein B sequence: PSRDRKETSI
Protein B construct: biotin-ttds-PSRDRKETSI

Average holdup BI: 0.57
Holdup BI standard deviation: 0.1
Normalized holdup BI: 0.65
Normalized holdup BI standard deviation: 0.11
Immobilized partner concentration in holdup experiment (10-6M): 28
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: 4.87


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: RALBP1
Protein B fragment: 646-655
Protein B site: PBM
Protein B sequence: PSRDRKETSI
Protein B construct: biotin-ttds-PSRDRKETSI

Average holdup BI: 0.38
Normalized holdup BI: 0.5
Immobilized partner concentration in holdup experiment (10-6M): 28
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: 4.58


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: RALBP1
Protein B fragment: 646-655
Protein B site: PBM
Protein B sequence: PSRDRKETSI
Protein B construct: biotin-ttds-PSRDRKETSI

Average holdup BI: 0.17
Normalized holdup BI: 0.2
Immobilized partner concentration in holdup experiment (10-6M): 28
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: 3.97


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: RALBP1
Protein B fragment: 646-655
Protein B site: PBM
Protein B sequence: PSRDRKETSI
Protein B construct: biotin-ttds-PSRDRKETSI

Average holdup BI: 0.61
Normalized holdup BI: 0.72
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: 5.23


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: RALBP1
Protein B fragment: 646-655
Protein B site: PBM
Protein B sequence: PSRDRKETSI
Protein B construct: biotin-ttds-PSRDRKETSI

Average holdup BI: 0.4
Normalized holdup BI: 0.5
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: 4.79


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: RALBP1
Protein B fragment: 646-655
Protein B site: PBM
Protein B sequence: PSRDRKETSI
Protein B construct: biotin-ttds-PSRDRKETSI

Average holdup BI: 0.15
Normalized holdup BI: 0.18
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: 4.1