Interaction Details


Details of experiments between Q5T2W1 and P27228
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: HPV35-E6
Fragment: 140-149
Site: PBM
Sequence: SKPTRRETEV
Construct: biotin-ttds-SKPTRRETEV
Average holdup BI: 0.15
Holdup BI standard deviation: 0.02
Normalized holdup BI: 0.16
Normalized holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 18.55
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: 4.04
PUBMED ID: 36115835

Experiment series 2

Protein A

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

Protein B

Protein: HPV35-E6
Fragment: 140-149
Site: PBM
Sequence: SKPTRRETEV
Construct: biotin-ttds-SKPTRRETEV
Average holdup BI: 0.25
Immobilized partner concentration (10-6M): 39.7
Experiment method: CALIP_holdup
Details of affinity fitting: BSA and lysozyme internal standards + bacterial cell lysate + normal layout
Number of measurements: 1
Measured pKd: 3.93
PUBMED ID: 36115835

Experiment series 3

Protein A

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

Protein B

Protein: HPV35-E6
Fragment: 140-149
Site: PBM
Sequence: SKPTRRETEV
Construct: biotin-ttds-SKPTRRETEV
Measured dissociation constant (10-6M): 146.23
Standard deviation of dissociation constant (10-6M): 69.77
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
Number of measurements: 3
Measured pKd: 3.83
PUBMED ID: 36115835

Experiment series 4

Protein A

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

Protein B

Protein: HPV35-E6
Fragment: 140-149
Site: PBM
Sequence: SKPTRRETEV
Construct: biotin-ttds-SKPTRRETEV
Average holdup BI: 0.07
Normalized holdup BI: 0.07
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: 3.62
PUBMED ID: 36115835
Experimental Details

not reported internal standards of holdup layout #6

Experiment series 5

Protein A

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

Protein B

Protein: HPV35-E6
Fragment: 140-149
Site: PBM
Sequence: SKPTRRETEV
Construct: biotin-ttds-SKPTRRETEV
Average holdup BI: 0.06
Normalized holdup BI: 0.06
Immobilized partner concentration (10-6M): 18.55
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: 3.57
Experimental Details

not reported internal standards of holdup layout #5

Experiment series 6

Protein A

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

Protein B

Protein: HPV35-E6
Fragment: 140-149
Site: PBM
Sequence: SKPTRRETEV
Construct: biotin-ttds-SKPTRRETEV
Average holdup BI: 0.05
Immobilized partner concentration (10-6M): 39.7
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: HPV35-E6
Fragment: 140-149
Site: PBM
Sequence: SKPTRRETEV
Construct: biotin-ttds-SKPTRRETEV
Average holdup BI: -0.06
Normalized holdup BI: -0.06
Immobilized partner concentration (10-6M): 18.55
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 8

Protein A

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

Protein B

Protein: HPV35-E6
Fragment: 140-149
Site: PBM
Sequence: SKPTRRETEV
Construct: biotin-ttds-SKPTRRETEV
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

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: HPV35-E6
Fragment: 140-149
Site: PBM
Sequence: SKPTRRETEV
Construct: biotin-ttds-SKPTRRETEV
Average holdup BI: -0.03
Normalized holdup BI: -0.03
Immobilized partner concentration (10-6M): 18.55
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 10

Protein A

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

Protein B

Protein: HPV35-E6
Fragment: 140-149
Site: PBM
Sequence: SKPTRRETEV
Construct: biotin-ttds-SKPTRRETEV
Average holdup BI: 0.08
Immobilized partner concentration (10-6M): 39.7
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: HPV35-E6
Fragment: 140-149
Site: PBM
Sequence: SKPTRRETEV
Construct: biotin-ttds-SKPTRRETEV
Average holdup BI: -0.04
Normalized holdup BI: -0.04
Immobilized partner concentration (10-6M): 18.55
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 12

Protein A

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

Protein B

Protein: HPV35-E6
Fragment: 140-149
Site: PBM
Sequence: SKPTRRETEV
Construct: biotin-ttds-SKPTRRETEV
Average holdup BI: 0.01
Normalized holdup BI: 0.01
Immobilized partner concentration (10-6M): 18.55
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 13

Protein A

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

Protein B

Protein: HPV35-E6
Fragment: 140-149
Site: PBM
Sequence: SKPTRRETEV
Construct: biotin-ttds-SKPTRRETEV
Average holdup BI: 0.02
Immobilized partner concentration (10-6M): 39.7
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: HPV35-E6
Fragment: 140-149
Site: PBM
Sequence: SKPTRRETEV
Construct: biotin-ttds-SKPTRRETEV
Average holdup BI: 0.05
Normalized holdup BI: 0.05
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

not reported internal standards of holdup layout #6

Experiment series 15

Protein A

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

Protein B

Protein: HPV35-E6
Fragment: 140-149
Site: PBM
Modification: (TPO)147
Sequence: SKPTRRETEV
Construct: Biotin-ttds-SKPTRREpTEV
Average holdup BI: -0.03
Normalized holdup BI: -0.04
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

Experiment series 16

Protein A

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

Protein B

Protein: HPV35-E6
Fragment: 140-149
Site: PBM
Modification: (TPO)147
Sequence: SKPTRRETEV
Construct: Biotin-ttds-SKPTRREpTEV
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

Experiment series 17

Protein A

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

Protein B

Protein: HPV35-E6
Fragment: 140-149
Site: PBM
Modification: (TPO)147
Sequence: SKPTRRETEV
Construct: Biotin-ttds-SKPTRREpTEV
Average holdup BI: 0
Normalized holdup BI: 0
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