Interaction Details

Details of experiments between Q86UT5 and Q15418
Note that the results of all experiments are listed, regardless of the modification states of the fragments.

Experiment series 1

Protein A

Protein: NHERF4
Fragment: 106-208
Site: PDZ_1
Construct: his6-MBP-TEVsite-PDZ

Protein B

Protein: RPS6KA1
Fragment: 726-735
Site: PBM
Sequence: RVRKLPSTTL
Construct: Biotin-ado-ado-RVRKLPSTTL

Average holdup BI: 0.11

Normalized holdup BI: 0.11

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: 3.85

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: NHERF4
Fragment: 106-208
Site: PDZ_1
Construct: his6-MBP-TEVsite-PDZ

Protein B

Protein: RPS6KA1
Fragment: 725-735
Site: PBM
Sequence: RVRKLPSTTL
Construct: biotin-ttds-RRVRKLPSTTL

Average holdup BI: 0.06

Normalized holdup BI: 0.06

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: 3.57

PUBMED ID: 36115835

Experiment series 3

Protein A

Protein: NHERF4
Fragment: 464-550
Site: PDZ_4
Construct: his6-MBP-TEVsite-PDZ

Protein B

Protein: RPS6KA1
Fragment: 725-735
Site: PBM
Sequence: RVRKLPSTTL
Construct: biotin-ttds-RRVRKLPSTTL

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: 3.49

PUBMED ID: 36115835

Experiment series 4

Protein A

Protein: NHERF4
Fragment: 106-208
Site: PDZ_1
Construct: his6-MBP-TEVsite-PDZ

Protein B

Protein: RPS6KA1
Fragment: 725-735
Site: PBM
Sequence: RVRKLPSTTL
Construct: biotin-ttds-RRVRKLPSTTL

Average holdup BI: 0.05

Holdup BI standard deviation: 0.04

Immobilized partner concentration (10^-6M): 31

Experiment method: CALIP_holdup

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

Number of measurements: 4

The affinity was below the detection threshold of the assay.

PUBMED ID: 36115835

Experiment series 5

Protein A

Protein: NHERF4
Fragment: 220-307
Site: PDZ_2
Construct: his6-MBP-TEVsite-PDZ

Protein B

Protein: RPS6KA1
Fragment: 725-735
Site: PBM
Sequence: RVRKLPSTTL
Construct: biotin-ttds-RRVRKLPSTTL

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

Experiment series 6

Protein A

Protein: NHERF4
Fragment: 220-307
Site: PDZ_2
Construct: his6-MBP-TEVsite-PDZ

Protein B

Protein: RPS6KA1
Fragment: 725-735
Site: PBM
Sequence: RVRKLPSTTL
Construct: biotin-ttds-RRVRKLPSTTL

Average holdup BI: -0.02

Holdup BI standard deviation: 0.02

Immobilized partner concentration (10^-6M): 31

Experiment method: CALIP_holdup

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

Number of measurements: 4

The affinity was below the detection threshold of the assay.

PUBMED ID: 36115835

Experiment series 7

Protein A

Protein: NHERF4
Fragment: 220-307
Site: PDZ_2
Construct: his6-MBP-TEVsite-PDZ

Protein B

Protein: RPS6KA1
Fragment: 726-735
Site: PBM
Sequence: RVRKLPSTTL
Construct: Biotin-ado-ado-RVRKLPSTTL

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

Experiment series 8

Protein A

Protein: NHERF4
Fragment: 325-412
Site: PDZ_3
Construct: his6-MBP-TEVsite-PDZ

Protein B

Protein: RPS6KA1
Fragment: 725-735
Site: PBM
Sequence: RVRKLPSTTL
Construct: biotin-ttds-RRVRKLPSTTL

Average holdup BI: 0.03

Holdup BI standard deviation: 0.05

Immobilized partner concentration (10^-6M): 31

Experiment method: CALIP_holdup

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

Number of measurements: 4

The affinity was below the detection threshold of the assay.

PUBMED ID: 36115835

Experiment series 9

Protein A

Protein: NHERF4
Fragment: 464-550
Site: PDZ_4
Construct: his6-MBP-TEVsite-PDZ

Protein B

Protein: RPS6KA1
Fragment: 725-735
Site: PBM
Sequence: RVRKLPSTTL
Construct: biotin-ttds-RRVRKLPSTTL

Average holdup BI: 0.05

Holdup BI standard deviation: 0.05

Immobilized partner concentration (10^-6M): 31

Experiment method: CALIP_holdup

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

Number of measurements: 4

The affinity was below the detection threshold of the assay.

PUBMED ID: 36115835

Experiment series 10

Protein A

Protein: NHERF4
Fragment: 464-550
Site: PDZ_4
Construct: his6-MBP-TEVsite-PDZ

Protein B

Protein: RPS6KA1
Fragment: 726-735
Site: PBM
Sequence: RVRKLPSTTL
Construct: Biotin-ado-ado-RVRKLPSTTL

Average holdup BI: -0.08

Normalized holdup BI: -0.08

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 11

Protein A

Protein: NHERF4
Fragment: 106-208
Site: PDZ_1
Construct: his6-MBP-TEVsite-PDZ

Protein B

Protein: RPS6KA1
Fragment: 725-735
Site: PBM
Modification: (SEP)732
Sequence: RVRKLPSTTL
Construct: biotin-ttds-RRVRKLPpSTTL

Average holdup BI: 0.09

Normalized holdup BI: 0.09

Immobilized partner concentration (10^-6M): 15

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: 3.83

Experimental Details

not reported internal standards of holdup layout #5

Experiment series 12

Protein A

Protein: NHERF4
Fragment: 464-550
Site: PDZ_4
Construct: his6-MBP-TEVsite-PDZ

Protein B

Protein: RPS6KA1
Fragment: 725-735
Site: PBM
Modification: (SEP)732
Sequence: RVRKLPSTTL
Construct: biotin-ttds-RRVRKLPpSTTL

Average holdup BI: 0.06

Normalized holdup BI: 0.06

Immobilized partner concentration (10^-6M): 15

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: 3.63

Experimental Details

not reported internal standards of holdup layout #5

Experiment series 13

Protein A

Protein: NHERF4
Fragment: 106-208
Site: PDZ_1
Construct: his6-MBP-TEVsite-PDZ

Protein B

Protein: RPS6KA1
Fragment: 725-735
Site: PBM
Modification: (SEP)732
Sequence: RVRKLPSTTL
Construct: biotin-ttds-RRVRKLPpSTTL

Average holdup BI: -0.02

Normalized holdup BI: -0.02

Immobilized partner concentration (10^-6M): 15

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 14

Protein A

Protein: NHERF4
Fragment: 106-208
Site: PDZ_1
Construct: his6-MBP-TEVsite-PDZ

Protein B

Protein: RPS6KA1
Fragment: 725-735
Site: PBM
Modification: (SEP)732
Sequence: RVRKLPSTTL
Construct: biotin-ttds-RRVRKLPpSTTL

Average holdup BI: 0.02

Holdup BI standard deviation: 0.08

Immobilized partner concentration (10^-6M): 40

Experiment method: CALIP_holdup

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

Number of measurements: 3

The affinity was below the detection threshold of the assay.

PUBMED ID: 36115835

Experiment series 15

Protein A

Protein: NHERF4
Fragment: 220-307
Site: PDZ_2
Construct: his6-MBP-TEVsite-PDZ

Protein B

Protein: RPS6KA1
Fragment: 725-735
Site: PBM
Modification: (SEP)732
Sequence: RVRKLPSTTL
Construct: biotin-ttds-RRVRKLPpSTTL

Average holdup BI: 0

Normalized holdup BI: 0

Immobilized partner concentration (10^-6M): 15

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: NHERF4
Fragment: 220-307
Site: PDZ_2
Construct: his6-MBP-TEVsite-PDZ

Protein B

Protein: RPS6KA1
Fragment: 725-735
Site: PBM
Modification: (SEP)732
Sequence: RVRKLPSTTL
Construct: biotin-ttds-RRVRKLPpSTTL

Average holdup BI: 0.02

Normalized holdup BI: 0.02

Immobilized partner concentration (10^-6M): 15

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 17

Protein A

Protein: NHERF4
Fragment: 220-307
Site: PDZ_2
Construct: his6-MBP-TEVsite-PDZ

Protein B

Protein: RPS6KA1
Fragment: 725-735
Site: PBM
Modification: (SEP)732
Sequence: RVRKLPSTTL
Construct: biotin-ttds-RRVRKLPpSTTL

Average holdup BI: -0.01

Holdup BI standard deviation: 0.03

Immobilized partner concentration (10^-6M): 40

Experiment method: CALIP_holdup

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

Number of measurements: 3

The affinity was below the detection threshold of the assay.

PUBMED ID: 36115835

Experiment series 18

Protein A

Protein: NHERF4
Fragment: 325-412
Site: PDZ_3
Construct: his6-MBP-TEVsite-PDZ

Protein B

Protein: RPS6KA1
Fragment: 725-735
Site: PBM
Modification: (SEP)732
Sequence: RVRKLPSTTL
Construct: biotin-ttds-RRVRKLPpSTTL

Average holdup BI: 0.04

Holdup BI standard deviation: 0.08

Immobilized partner concentration (10^-6M): 40

Experiment method: CALIP_holdup

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

Number of measurements: 3

The affinity was below the detection threshold of the assay.

PUBMED ID: 36115835

Experiment series 19

Protein A

Protein: NHERF4
Fragment: 464-550
Site: PDZ_4
Construct: his6-MBP-TEVsite-PDZ

Protein B

Protein: RPS6KA1
Fragment: 725-735
Site: PBM
Modification: (SEP)732
Sequence: RVRKLPSTTL
Construct: biotin-ttds-RRVRKLPpSTTL

Average holdup BI: -0

Normalized holdup BI: -0

Immobilized partner concentration (10^-6M): 15

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 20

Protein A

Protein: NHERF4
Fragment: 464-550
Site: PDZ_4
Construct: his6-MBP-TEVsite-PDZ

Protein B

Protein: RPS6KA1
Fragment: 725-735
Site: PBM
Modification: (SEP)732
Sequence: RVRKLPSTTL
Construct: biotin-ttds-RRVRKLPpSTTL

Average holdup BI: -0

Holdup BI standard deviation: 0.08

Immobilized partner concentration (10^-6M): 40

Experiment method: CALIP_holdup

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

Number of measurements: 3

The affinity was below the detection threshold of the assay.

PUBMED ID: 36115835

Experiment series 21

Protein A

Protein: NHERF4
Fragment: 106-208
Site: PDZ_1
Construct: his6-MBP-TEVsite-PDZ

Protein B

Protein: RPS6KA1
Fragment: 725-735
Site: PBM
Modification: (TPO)733
Sequence: RVRKLPSTTL
Construct: biotin-ttds-RRVRKLPSpTTL

Average holdup BI: 0.08

Normalized holdup BI: 0.08

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: 3.69

PUBMED ID: 36115835

Experiment series 22

Protein A

Protein: NHERF4
Fragment: 220-307
Site: PDZ_2
Construct: his6-MBP-TEVsite-PDZ

Protein B

Protein: RPS6KA1
Fragment: 725-735
Site: PBM
Modification: (TPO)733
Sequence: RVRKLPSTTL
Construct: biotin-ttds-RRVRKLPSpTTL

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

Experiment series 23

Protein A

Protein: NHERF4
Fragment: 464-550
Site: PDZ_4
Construct: his6-MBP-TEVsite-PDZ

Protein B

Protein: RPS6KA1
Fragment: 725-735
Site: PBM
Modification: (TPO)733
Sequence: RVRKLPSTTL
Construct: biotin-ttds-RRVRKLPSpTTL

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

Experiment series 24

Protein A

Protein: NHERF4
Fragment: 106-208
Site: PDZ_1
Construct: his6-MBP-TEVsite-PDZ

Protein B

Protein: RPS6KA1
Fragment: 725-735
Site: PBM
Modification: (TPO)734
Sequence: RVRKLPSTTL
Construct: Biotin-ttds-RRVRKLPSTpTL

Average holdup BI: 0.29

Normalized holdup BI: 0.29

Immobilized partner concentration (10^-6M): 25

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: 4.23

PUBMED ID: 36115835

Experiment series 25

Protein A

Protein: NHERF4
Fragment: 220-307
Site: PDZ_2
Construct: his6-MBP-TEVsite-PDZ

Protein B

Protein: RPS6KA1
Fragment: 725-735
Site: PBM
Modification: (TPO)734
Sequence: RVRKLPSTTL
Construct: Biotin-ttds-RRVRKLPSTpTL

Average holdup BI: 0.02

Normalized holdup BI: 0.02

Immobilized partner concentration (10^-6M): 25

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: NHERF4
Fragment: 464-550
Site: PDZ_4
Construct: his6-MBP-TEVsite-PDZ

Protein B

Protein: RPS6KA1
Fragment: 725-735
Site: PBM
Modification: (TPO)734
Sequence: RVRKLPSTTL
Construct: Biotin-ttds-RRVRKLPSTpTL

Average holdup BI: 0.05

Normalized holdup BI: 0.05

Immobilized partner concentration (10^-6M): 25

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