Details of experiments between P78352 and Q7Z628
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: 155-249
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ

Protein B protein: NET1
Protein B fragment: 587-596
Protein B site: PBM
Protein B sequence: SGGKRKETLV
Protein B construct: biotin-ttds-SGGKRKETLV

Average holdup BI: 0.92
Immobilized partner concentration in holdup experiment (10-6M): 35
Experiment method: CALIP_holdup
Details of affinity fitting: BSA and lysozyme internal standards + bacterial cell lysate + normal layout
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 5.56


Experiment series 2

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

Protein B protein: NET1
Protein B fragment: 587-596
Protein B site: PBM
Protein B sequence: SGGKRKETLV
Protein B construct: biotin-ttds-SGGKRKETLV

Average holdup BI: 0.78
Immobilized partner concentration in holdup experiment (10-6M): 35
Experiment method: CALIP_holdup
Details of affinity fitting: BSA and lysozyme internal standards + bacterial cell lysate + normal layout
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 5.04


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: NET1
Protein B fragment: 587-596
Protein B site: PBM
Protein B sequence: SGGKRKETLV
Protein B construct: biotin-ttds-SGGKRKETLV

Average holdup BI: 0.57
Immobilized partner concentration in holdup experiment (10-6M): 35
Experiment method: CALIP_holdup
Details of affinity fitting: BSA and lysozyme internal standards + bacterial cell lysate + normal layout
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 4.61


Experiment series 4

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

Protein B protein: NET1
Protein B fragment: 587-596
Protein B site: PBM
Protein B sequence: SGGKRKETLV
Protein B construct: biotin-ttds-SGGKRKETLV

Measured dissociation constant (10-6M): 10.63
Standard deviation of dissociation constant (10-6M): 0.47
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit

PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 4.97


Experiment series 5

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

Protein B protein: NET1
Protein B fragment: 587-596
Protein B site: PBM
Protein B sequence: SGGKRKETLV
Protein B construct: biotin-ttds-SGGKRKETLV

Average holdup BI: 0.69
Holdup BI standard deviation: 0.01
Normalized holdup BI: 0.89
Normalized holdup BI standard deviation: 0.02
Immobilized partner concentration in holdup experiment (10-6M): 17.9
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: 5.77


Experiment series 6

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

Protein B protein: NET1
Protein B fragment: 587-596
Protein B site: PBM
Protein B sequence: SGGKRKETLV
Protein B construct: biotin-ttds-SGGKRKETLV

Average holdup BI: 0.59
Holdup BI standard deviation: 0
Normalized holdup BI: 0.71
Normalized holdup BI standard deviation: 0.01
Immobilized partner concentration in holdup experiment (10-6M): 17.9
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: 5.21


Experiment series 7

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

Protein B protein: NET1
Protein B fragment: 587-596
Protein B site: PBM
Protein B sequence: SGGKRKETLV
Protein B construct: biotin-ttds-SGGKRKETLV

Average holdup BI: 0.49
Holdup BI standard deviation: 0.09
Normalized holdup BI: 0.56
Normalized holdup BI standard deviation: 0.1
Immobilized partner concentration in holdup experiment (10-6M): 17.9
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: 3
Measured pKd: 4.92


Experiment series 8

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

Protein B protein: NET1
Protein B fragment: 587-596
Protein B site: PBM
Protein B sequence: SGGKRKETLV
Protein B construct: biotin-ttds-SGGKRKETLV

Average holdup BI: 0.55
Normalized holdup BI: 0.55
Immobilized partner concentration in holdup experiment (10-6M): 17.9
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 #5
Number of measurements: 1
Measured pKd: 4.9


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: NET1
Protein B fragment: 587-596
Protein B site: PBM
Protein B sequence: SGGKRKETLV
Protein B construct: biotin-ttds-SGGKRKETLV

Average holdup BI: 0.68
Normalized holdup BI: 0.82
Immobilized partner concentration in holdup experiment (10-6M): 17.9
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 #5
Number of measurements: 1
Measured pKd: 5.49


Experiment series 10

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

Protein B protein: NET1
Protein B fragment: 587-596
Protein B site: PBM
Protein B sequence: SGGKRKETLV
Protein B construct: biotin-ttds-SGGKRKETLV

Average holdup BI: 0.37
Normalized holdup BI: 0.46
Immobilized partner concentration in holdup experiment (10-6M): 17.9
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 #5
Number of measurements: 1
Measured pKd: 4.72


Experiment series 11

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

Protein B protein: NET1
Protein B fragment: 587-596
Protein B site: PBM
Protein B sequence: SGGKRKETLV
Protein B construct: biotin-ttds-SGGKRKETLV

Average holdup BI: 0.38
Normalized holdup BI: 0.43
Immobilized partner concentration in holdup experiment (10-6M): 17.9
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 #5
Number of measurements: 1
Measured pKd: 4.67


Experiment series 12

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

Protein B protein: NET1
Protein B fragment: 587-596
Protein B site: PBM
Protein B sequence: SGGKRKETLV
Protein B construct: Biotin-ado-ado-SGGKRKETLV

Average holdup BI: 0.67
Normalized holdup BI: 0.79
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: 5.39


Experiment series 13

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

Protein B protein: NET1
Protein B fragment: 587-596
Protein B site: PBM
Protein B sequence: SGGKRKETLV
Protein B construct: Biotin-ado-ado-SGGKRKETLV

Average holdup BI: 0.78
Normalized holdup BI: 0.97
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: 6.36


Experiment series 14

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

Protein B protein: NET1
Protein B fragment: 587-596
Protein B site: PBM
Protein B sequence: SGGKRKETLV
Protein B construct: Biotin-ado-ado-SGGKRKETLV

Average holdup BI: 0.67
Normalized holdup BI: 0.79
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: 5.4