Interaction Details


Details of experiments between Q5T2W1 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: NHERF3
Fragment: 2-107
Site: PDZ_1
Construct: his6-MBP-TEVsite-PDZ

Protein B

Protein: NET1
Fragment: 587-596
Site: PBM
Sequence: SGGKRKETLV
Construct: Biotin-ado-ado-SGGKRKETLV
Average holdup BI: 0.41
Normalized holdup BI: 0.44
Immobilized partner concentration (10-6M): 18
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
Number of measurements: 1
Measured pKd: 4.68
PUBMED ID: 36115835
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

Experiment series 2

Protein A

Protein: NHERF3
Fragment: 2-107
Site: PDZ_1
Construct: his6-MBP-TEVsite-PDZ

Protein B

Protein: NET1
Fragment: 587-596
Site: PBM
Sequence: SGGKRKETLV
Construct: biotin-ttds-SGGKRKETLV
Average holdup BI: 0.47
Immobilized partner concentration (10-6M): 35
Experiment method: CALIP_holdup
Details of affinity fitting: BSA and lysozyme internal standards + bacterial cell lysate + normal layout
Number of measurements: 1
Measured pKd: 4.42
PUBMED ID: 36115835

Experiment series 3

Protein A

Protein: NHERF3
Fragment: 2-107
Site: PDZ_1
Construct: his6-MBP-TEVsite-PDZ

Protein B

Protein: NET1
Fragment: 587-596
Site: PBM
Sequence: SGGKRKETLV
Construct: biotin-ttds-SGGKRKETLV
Average holdup BI: 0.26
Holdup BI standard deviation: 0.01
Normalized holdup BI: 0.27
Normalized holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 17.9
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
Number of measurements: 2
Measured pKd: 4.35
PUBMED ID: 36115835

Experiment series 4

Protein A

Protein: NHERF3
Fragment: 2-107
Site: PDZ_1
Construct: his6-MBP-TEVsite-PDZ

Protein B

Protein: NET1
Fragment: 587-596
Site: PBM
Sequence: SGGKRKETLV
Construct: biotin-ttds-SGGKRKETLV
Average holdup BI: 0.21
Normalized holdup BI: 0.23
Immobilized partner concentration (10-6M): 17.9
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
Number of measurements: 1
Measured pKd: 4.25
Experimental Details

not reported internal standards of holdup layout #5

Experiment series 5

Protein A

Protein: NHERF3
Fragment: 2-107
Site: PDZ_1
Construct: his6-MBP-TEVsite-PDZ

Protein B

Protein: NET1
Fragment: 587-596
Site: PBM
Sequence: SGGKRKETLV
Construct: biotin-ttds-SGGKRKETLV
Measured dissociation constant (10-6M): 73.04
Standard deviation of dissociation constant (10-6M): 26.23
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
Number of measurements: 3
Measured pKd: 4.14
PUBMED ID: 36115835

Experiment series 6

Protein A

Protein: NHERF3
Fragment: 132-224
Site: PDZ_2
Construct: his6-MBP-TEVsite-PDZ

Protein B

Protein: NET1
Fragment: 587-596
Site: PBM
Sequence: SGGKRKETLV
Construct: biotin-ttds-SGGKRKETLV
Average holdup BI: -0.01
Immobilized partner concentration (10-6M): 35
Experiment method: CALIP_holdup
Details of affinity fitting: BSA and lysozyme internal standards + bacterial cell lysate + normal layout
Number of measurements: 1
The affinity was below the detection threshold of the assay.
PUBMED ID: 36115835

Experiment series 7

Protein A

Protein: NHERF3
Fragment: 238-329
Site: PDZ_3
Construct: his6-MBP-TEVsite-PDZ

Protein B

Protein: NET1
Fragment: 587-596
Site: PBM
Sequence: SGGKRKETLV
Construct: biotin-ttds-SGGKRKETLV
Average holdup BI: -0.03
Normalized holdup BI: -0.03
Immobilized partner concentration (10-6M): 17.9
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experimental Details

not reported internal standards of holdup layout #5

Experiment series 8

Protein A

Protein: NHERF3
Fragment: 238-329
Site: PDZ_3
Construct: his6-MBP-TEVsite-PDZ

Protein B

Protein: NET1
Fragment: 587-596
Site: PBM
Sequence: SGGKRKETLV
Construct: Biotin-ado-ado-SGGKRKETLV
Average holdup BI: -0.03
Normalized holdup BI: -0.03
Immobilized partner concentration (10-6M): 18
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
Number of measurements: 1
The affinity was below the detection threshold of the assay.
PUBMED ID: 36115835
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

Experiment series 9

Protein A

Protein: NHERF3
Fragment: 238-329
Site: PDZ_3
Construct: his6-MBP-TEVsite-PDZ

Protein B

Protein: NET1
Fragment: 587-596
Site: PBM
Sequence: SGGKRKETLV
Construct: biotin-ttds-SGGKRKETLV
Average holdup BI: 0
Normalized holdup BI: 0
Immobilized partner concentration (10-6M): 17.9
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
Number of measurements: 1
The affinity was below the detection threshold of the assay.
PUBMED ID: 36115835

Experiment series 10

Protein A

Protein: NHERF3
Fragment: 238-329
Site: PDZ_3
Construct: his6-MBP-TEVsite-PDZ

Protein B

Protein: NET1
Fragment: 587-596
Site: PBM
Sequence: SGGKRKETLV
Construct: biotin-ttds-SGGKRKETLV
Average holdup BI: 0.05
Immobilized partner concentration (10-6M): 35
Experiment method: CALIP_holdup
Details of affinity fitting: BSA and lysozyme internal standards + bacterial cell lysate + normal layout
Number of measurements: 1
The affinity was below the detection threshold of the assay.
PUBMED ID: 36115835

Experiment series 11

Protein A

Protein: NHERF3
Fragment: 375-459
Site: PDZ_4
Construct: his6-MBP-TEVsite-PDZ

Protein B

Protein: NET1
Fragment: 587-596
Site: PBM
Sequence: SGGKRKETLV
Construct: biotin-ttds-SGGKRKETLV
Average holdup BI: -0.1
Normalized holdup BI: -0.1
Immobilized partner concentration (10-6M): 17.9
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experimental Details

not reported internal standards of holdup layout #5

Experiment series 12

Protein A

Protein: NHERF3
Fragment: 375-459
Site: PDZ_4
Construct: his6-MBP-TEVsite-PDZ

Protein B

Protein: NET1
Fragment: 587-596
Site: PBM
Sequence: SGGKRKETLV
Construct: biotin-ttds-SGGKRKETLV
Average holdup BI: -0.01
Normalized holdup BI: -0.01
Immobilized partner concentration (10-6M): 17.9
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
Number of measurements: 1
The affinity was below the detection threshold of the assay.
PUBMED ID: 36115835

Experiment series 13

Protein A

Protein: NHERF3
Fragment: 375-459
Site: PDZ_4
Construct: his6-MBP-TEVsite-PDZ

Protein B

Protein: NET1
Fragment: 587-596
Site: PBM
Sequence: SGGKRKETLV
Construct: biotin-ttds-SGGKRKETLV
Average holdup BI: 0
Immobilized partner concentration (10-6M): 35
Experiment method: CALIP_holdup
Details of affinity fitting: BSA and lysozyme internal standards + bacterial cell lysate + normal layout
Number of measurements: 1
The affinity was below the detection threshold of the assay.
PUBMED ID: 36115835

Experiment series 14

Protein A

Protein: NHERF3
Fragment: 375-459
Site: PDZ_4
Construct: his6-MBP-TEVsite-PDZ

Protein B

Protein: NET1
Fragment: 587-596
Site: PBM
Sequence: SGGKRKETLV
Construct: Biotin-ado-ado-SGGKRKETLV
Average holdup BI: 0.01
Normalized holdup BI: 0.01
Immobilized partner concentration (10-6M): 18
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
Number of measurements: 1
The affinity was below the detection threshold of the assay.
PUBMED ID: 36115835
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