Interaction Details

Details of experiments between Q5T2W1 and P60484
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: PTEN
Fragment: 391-403
Site: PBM
Sequence: EDQHTQITKV
Construct: Biotin-ttds-PFDEDQHTQITKV

Average holdup BI: 0.16

Immobilized partner concentration (10^-6M): 17

Experiment method: CALIP_holdup

Details of affinity fitting: lysozyme internal standard

Number of measurements: 1

Measured pK_d: 4.07

PUBMED ID: 33382810

Experiment series 2

Protein A

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

Protein B

Protein: PTEN
Fragment: 394-403
Site: PBM
Sequence: EDQHTQITKV
Construct: Biotin-ado-ado-EDQHTQITKV

Average holdup BI: 0.15

Normalized holdup BI: 0.16

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 pK_d: 4.04

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 3

Protein A

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

Protein B

Protein: PTEN
Fragment: 393-403
Site: PBM
Sequence: EDQHTQITKV
Construct: biotin-ttds-DEDQHTQITKV

Average holdup BI: 0.15

Normalized holdup BI: 0.16

Immobilized partner concentration (10^-6M): 19

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 pK_d: 4.03

PUBMED ID: 36115835

Experiment series 4

Protein A

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

Protein B

Protein: PTEN
Fragment: 393-403
Site: PBM
Sequence: EDQHTQITKV
Construct: biotin-ttds-DEDQHTQITKV

Average holdup BI: 0.15

Immobilized partner concentration (10^-6M): 36

Experiment method: CALIP_holdup

Details of affinity fitting: BSA and lysozyme internal standards + bacterial cell lysate + normal layout

Number of measurements: 1

Measured pK_d: 3.71

PUBMED ID: 36115835

Experiment series 5

Protein A

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

Protein B

Protein: PTEN
Fragment: 393-403
Site: PBM
Sequence: EDQHTQITKV
Construct: biotin-ttds-DEDQHTQITKV

Average holdup BI: 0.08

Normalized holdup BI: 0.08

Immobilized partner concentration (10^-6M): 19

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 pK_d: 3.67

PUBMED ID: 36115835

Experiment series 6

Protein A

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

Protein B

Protein: PTEN
Fragment: 393-403
Site: PBM
Sequence: EDQHTQITKV
Construct: biotin-ttds-DEDQHTQITKV

Average holdup BI: 0.13

Immobilized partner concentration (10^-6M): 36

Experiment method: CALIP_holdup

Details of affinity fitting: BSA and lysozyme internal standards + bacterial cell lysate + normal layout

Number of measurements: 1

Measured pK_d: 3.63

PUBMED ID: 36115835

Experiment series 7

Protein A

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

Protein B

Protein: PTEN
Fragment: 393-403
Site: PBM
Sequence: EDQHTQITKV
Construct: biotin-ttds-DEDQHTQITKV

Average holdup BI: 0.06

Normalized holdup BI: 0.06

Immobilized partner concentration (10^-6M): 19

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 pK_d: 3.53

PUBMED ID: 36115835

Experiment series 8

Protein A

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

Protein B

Protein: PTEN
Fragment: 394-403
Site: PBM
Sequence: EDQHTQITKV
Construct: Biotin-ado-ado-EDQHTQITKV

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

Measured pK_d: 3.48

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: 132-224
Site: PDZ_2
Construct: his6-MBP-TEVsite-PDZ

Protein B

Protein: PTEN
Fragment: 391-403
Site: PBM
Sequence: EDQHTQITKV
Construct: Biotin-ttds-PFDEDQHTQITKV

Average holdup BI: 0.06

Immobilized partner concentration (10^-6M): 17

Experiment method: CALIP_holdup

Details of affinity fitting: lysozyme internal standard

Number of measurements: 1

The affinity was below the detection threshold of the assay.

PUBMED ID: 33382810

Experiment series 10

Protein A

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

Protein B

Protein: PTEN
Fragment: 393-403
Site: PBM
Sequence: EDQHTQITKV
Construct: biotin-ttds-DEDQHTQITKV

Average holdup BI: 0.06

Immobilized partner concentration (10^-6M): 36

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: 2-107
Site: PDZ_1
Construct: his6-MBP-TEVsite-PDZ

Protein B

Protein: PTEN
Fragment: 393-403
Site: PBM
Sequence: EDQHTQITKV
Construct: biotin-ttds-DEDQHTQITKV

Measured dissociation constant (10^-6M): 0

Experiment method: competitive FP

Details of affinity fitting: Competitive binding equation in ProFit

Number of measurements: 3

The affinity was below the detection threshold of the assay.

PUBMED ID: 36115835

Experiment series 12

Protein A

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

Protein B

Protein: PTEN
Fragment: 391-403
Site: PBM
Sequence: EDQHTQITKV
Construct: Biotin-ttds-PFDEDQHTQITKV

Average holdup BI: 0.07

Immobilized partner concentration (10^-6M): 17

Experiment method: CALIP_holdup

Details of affinity fitting: lysozyme internal standard

Number of measurements: 1

The affinity was below the detection threshold of the assay.

PUBMED ID: 33382810

Experiment series 13

Protein A

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

Protein B

Protein: PTEN
Fragment: 391-403
Site: PBM
Sequence: EDQHTQITKV
Construct: Biotin-ttds-PFDEDQHTQITKV

Average holdup BI: -0.01

Immobilized partner concentration (10^-6M): 17

Experiment method: CALIP_holdup

Details of affinity fitting: lysozyme internal standard

Number of measurements: 1

The affinity was below the detection threshold of the assay.

PUBMED ID: 33382810

Experiment series 14

Protein A

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

Protein B

Protein: PTEN
Fragment: 393-403
Site: PBM
Sequence: EDQHTQITKV
Construct: biotin-ttds-DEDQHTQITKV

Average holdup BI: -0.01

Immobilized partner concentration (10^-6M): 36

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 15

Protein A

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

Protein B

Protein: PTEN
Fragment: 394-403
Site: PBM
Sequence: EDQHTQITKV
Construct: Biotin-ado-ado-EDQHTQITKV

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 16

Protein A

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

Protein B

Protein: PTEN
Fragment: 393-403
Site: PBM
Modification: (ALY)402
Sequence: EDQHTQITKV
Construct: biotin-ttds-DEDQHTQITKacV

Average holdup BI: 0.45

Immobilized partner concentration (10^-6M): 29

Experiment method: CALIP_holdup

Details of affinity fitting: BSA and lysozyme internal standards + bacterial cell lysate + normal layout

Number of measurements: 1

Measured pK_d: 4.48

PUBMED ID: 36115835

Experiment series 17

Protein A

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

Protein B

Protein: PTEN
Fragment: 393-403
Site: PBM
Modification: (ALY)402
Sequence: EDQHTQITKV
Construct: biotin-ttds-DEDQHTQITKacV

Average holdup BI: 0.18

Normalized holdup BI: 0.19

Immobilized partner concentration (10^-6M): 17

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 pK_d: 4.16

PUBMED ID: 36115835

Experiment series 18

Protein A

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

Protein B

Protein: PTEN
Fragment: 393-403
Site: PBM
Modification: (ALY)402
Sequence: EDQHTQITKV
Construct: biotin-ttds-DEDQHTQITKacV

Average holdup BI: 0.16

Normalized holdup BI: 0.17

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 pK_d: 4.08

PUBMED ID: 36115835

Experimental Details

not reported internal standards of holdup layout #6

Experiment series 19

Protein A

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

Protein B

Protein: PTEN
Fragment: 393-403
Site: PBM
Modification: (ALY)402
Sequence: EDQHTQITKV
Construct: biotin-ttds-DEDQHTQITKacV

Average holdup BI: 0.13

Normalized holdup BI: 0.14

Immobilized partner concentration (10^-6M): 17

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 pK_d: 3.99

Experimental Details

not reported internal standards of holdup layout #5

Experiment series 20

Protein A

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

Protein B

Protein: PTEN
Fragment: 393-403
Site: PBM
Modification: (ALY)402
Sequence: EDQHTQITKV
Construct: biotin-ttds-DEDQHTQITKacV

Average holdup BI: 0.11

Immobilized partner concentration (10^-6M): 29

Experiment method: CALIP_holdup

Details of affinity fitting: BSA and lysozyme internal standards + bacterial cell lysate + normal layout

Number of measurements: 1

Measured pK_d: 3.61

PUBMED ID: 36115835

Experiment series 21

Protein A

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

Protein B

Protein: PTEN
Fragment: 393-403
Site: PBM
Modification: (ALY)402
Sequence: EDQHTQITKV
Construct: biotin-ttds-DEDQHTQITKacV

Measured dissociation constant (10^-6M): 0

Experiment method: competitive FP

Details of affinity fitting: Competitive binding equation in ProFit

Number of measurements: 3

The affinity was below the detection threshold of the assay.

PUBMED ID: 36115835

Experiment series 22

Protein A

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

Protein B

Protein: PTEN
Fragment: 393-403
Site: PBM
Modification: (ALY)402
Sequence: EDQHTQITKV
Construct: biotin-ttds-DEDQHTQITKacV

Average holdup BI: -0.02

Normalized holdup BI: -0.02

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 23

Protein A

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

Protein B

Protein: PTEN
Fragment: 393-403
Site: PBM
Modification: (ALY)402
Sequence: EDQHTQITKV
Construct: biotin-ttds-DEDQHTQITKacV

Average holdup BI: 0.01

Normalized holdup BI: 0.01

Immobilized partner concentration (10^-6M): 17

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 24

Protein A

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

Protein B

Protein: PTEN
Fragment: 393-403
Site: PBM
Modification: (ALY)402
Sequence: EDQHTQITKV
Construct: biotin-ttds-DEDQHTQITKacV

Average holdup BI: 0.02

Immobilized partner concentration (10^-6M): 29

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 25

Protein A

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

Protein B

Protein: PTEN
Fragment: 393-403
Site: PBM
Modification: (ALY)402
Sequence: EDQHTQITKV
Construct: biotin-ttds-DEDQHTQITKacV

Average holdup BI: 0.05

Normalized holdup BI: 0.05

Immobilized partner concentration (10^-6M): 17

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 26

Protein A

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

Protein B

Protein: PTEN
Fragment: 393-403
Site: PBM
Modification: (ALY)402
Sequence: EDQHTQITKV
Construct: biotin-ttds-DEDQHTQITKacV

Average holdup BI: -0.02

Immobilized partner concentration (10^-6M): 29

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 27

Protein A

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

Protein B

Protein: PTEN
Fragment: 393-403
Site: PBM
Modification: (ALY)402
Sequence: EDQHTQITKV
Construct: biotin-ttds-DEDQHTQITKacV

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

not reported internal standards of holdup layout #6

Experiment series 28

Protein A

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

Protein B

Protein: PTEN
Fragment: 393-403
Site: PBM
Modification: (ALY)402
Sequence: EDQHTQITKV
Construct: biotin-ttds-DEDQHTQITKacV

Average holdup BI: 0.04

Normalized holdup BI: 0.04

Immobilized partner concentration (10^-6M): 17

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 29

Protein A

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

Protein B

Protein: PTEN
Fragment: 393-403
Site: PBM
Modification: (ALY)402
Sequence: EDQHTQITKV
Construct: biotin-ttds-DEDQHTQITKacV

Average holdup BI: 0.05

Normalized holdup BI: 0.05

Immobilized partner concentration (10^-6M): 17

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