Interaction Details


Details of experiments between Q14160 and P23634
Note that the results of all experiments are listed, regardless of the modification states of the fragments.


Experiment series 1

Protein A

Protein: SCRIB
Fragment: 1-1630
Construct: endogenous protein (Jurkat)

Protein B

Protein: ATP2B4
Fragment: 1232-1241
Site: PBM
Sequence: SSLQSLETSV
Construct: biotin-ttds-SSLQSLETSV
Experiment method: nHU-WB_Jurkat
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-08-03
Number of measurements: 3
Measured pKd: 5.91
PUBMED ID: 36542723

Experiment series 2

Protein A

Protein: SCRIB
Fragment: 718-814
Site: PDZ_1
Construct: his6-MBP-TEVsite-PDZ

Protein B

Protein: ATP2B4
Fragment: 1232-1241
Site: PBM
Sequence: SSLQSLETSV
Construct: biotin-ttds-SSLQSLETSV
Average holdup BI: 0.43
Immobilized partner concentration (10-6M): 7
Experiment method: SAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normal layout
Number of measurements: 1
Measured pKd: 5.15
PUBMED ID: 36115835

Experiment series 3

Protein A

Protein: SCRIB
Fragment: 999-1093
Site: PDZ_3
Construct: his6-MBP-TEVsite-PDZ

Protein B

Protein: ATP2B4
Fragment: 1232-1241
Site: PBM
Sequence: SSLQSLETSV
Construct: biotin-ttds-SSLQSLETSV
Average holdup BI: 0.6
Normalized holdup BI: 0.67
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: 5.11
PUBMED ID: 36115835

Experiment series 4

Protein A

Protein: SCRIB
Fragment: 999-1093
Site: PDZ_3
Construct: his6-MBP-TEVsite-PDZ

Protein B

Protein: ATP2B4
Fragment: 1232-1241
Site: PBM
Sequence: SSLQSLETSV
Construct: biotin-ttds-SSLQSLETSV
Average holdup BI: 0.56
Normalized holdup BI: 0.66
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: 5.1
PUBMED ID: 36115835
Experimental Details

not reported internal standards of holdup layout #6

Experiment series 5

Protein A

Protein: SCRIB
Fragment: 718-814
Site: PDZ_1
Construct: his6-MBP-TEVsite-PDZ

Protein B

Protein: ATP2B4
Fragment: 1232-1241
Site: PBM
Sequence: SSLQSLETSV
Construct: biotin-ttds-SSLQSLETSV
Average holdup BI: 0.66
Normalized holdup BI: 0.66
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: 5.1
PUBMED ID: 36115835
Experimental Details

not reported internal standards of holdup layout #6

Experiment series 6

Protein A

Protein: SCRIB
Fragment: 718-814
Site: PDZ_1
Construct: his6-MBP-TEVsite-PDZ

Protein B

Protein: ATP2B4
Fragment: 1232-1241
Site: PBM
Sequence: SSLQSLETSV
Construct: biotin-ttds-SSLQSLETSV
Average holdup BI: 0.59
Holdup BI standard deviation: 0.03
Normalized holdup BI: 0.62
Normalized holdup BI standard deviation: 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: 4
Measured pKd: 5.03
PUBMED ID: 36115835

Experiment series 7

Protein A

Protein: SCRIB
Fragment: 718-814
Site: PDZ_1
Construct: his6-MBP-TEVsite-PDZ

Protein B

Protein: ATP2B4
Fragment: 1232-1241
Site: PBM
Sequence: SSLQSLETSV
Construct: biotin-ttds-SSLQSLETSV
Measured dissociation constant (10-6M): 9.44
Standard deviation of dissociation constant (10-6M): 0.59
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
Number of measurements: 3
Measured pKd: 5.03
PUBMED ID: 36115835

Experiment series 8

Protein A

Protein: SCRIB
Fragment: 999-1093
Site: PDZ_3
Construct: his6-MBP-TEVsite-PDZ

Protein B

Protein: ATP2B4
Fragment: 1232-1241
Site: PBM
Sequence: SSLQSLETSV
Construct: biotin-ttds-SSLQSLETSV
Average holdup BI: 0.32
Immobilized partner concentration (10-6M): 7
Experiment method: SAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normal layout
Number of measurements: 1
Measured pKd: 4.92
PUBMED ID: 36115835

Experiment series 9

Protein A

Protein: SCRIB
Fragment: 858-949
Site: PDZ_2
Construct: his6-MBP-TEVsite-PDZ

Protein B

Protein: ATP2B4
Fragment: 1232-1241
Site: PBM
Sequence: SSLQSLETSV
Construct: biotin-ttds-SSLQSLETSV
Average holdup BI: 0.45
Normalized holdup BI: 0.56
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: 4.9
PUBMED ID: 36115835

Experiment series 10

Protein A

Protein: SCRIB
Fragment: 858-949
Site: PDZ_2
Construct: his6-MBP-TEVsite-PDZ

Protein B

Protein: ATP2B4
Fragment: 1232-1241
Site: PBM
Sequence: SSLQSLETSV
Construct: biotin-ttds-SSLQSLETSV
Average holdup BI: 0.55
Normalized holdup BI: 0.55
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: 4.89
PUBMED ID: 36115835
Experimental Details

not reported internal standards of holdup layout #6

Experiment series 11

Protein A

Protein: SCRIB
Fragment: 858-949
Site: PDZ_2
Construct: his6-MBP-TEVsite-PDZ

Protein B

Protein: ATP2B4
Fragment: 1232-1241
Site: PBM
Sequence: SSLQSLETSV
Construct: biotin-ttds-SSLQSLETSV
Average holdup BI: 0.16
Immobilized partner concentration (10-6M): 7
Experiment method: SAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normal layout
Number of measurements: 1
Measured pKd: 4.47
PUBMED ID: 36115835

Experiment series 12

Protein A

Protein: SCRIB
Fragment: 1090-1197
Site: PDZ_4
Construct: his6-MBP-TEVsite-PDZ

Protein B

Protein: ATP2B4
Fragment: 1232-1241
Site: PBM
Sequence: SSLQSLETSV
Construct: biotin-ttds-SSLQSLETSV
Average holdup BI: 0.13
Immobilized partner concentration (10-6M): 7
Experiment method: SAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normal layout
Number of measurements: 1
Measured pKd: 4.38
PUBMED ID: 36115835

Experiment series 13

Protein A

Protein: SCRIB
Fragment: 718-814
Site: PDZ_1
Construct: avi(biotin)-his6-MBP-TEVsite-PDZ

Protein B

Protein: ATP2B4
Fragment: 1-1241
Construct: endogenous protein (Jurkat)
Average holdup BI: 0.23
Immobilized partner concentration (10-6M): 10
P value: 2.19
Experiment method: nHU_Jurkat
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-08-03
Number of measurements: 3
The affinity was below the detection threshold of the assay.
PUBMED ID: 36542723
Experimental Details

-log10(P Two-sided unpaired T-test (6 vs 3))

Experiment series 14

Protein A

Protein: SCRIB
Fragment: 858-949
Site: PDZ_2
Construct: avi(biotin)-his6-MBP-TEVsite-PDZ

Protein B

Protein: ATP2B4
Fragment: 1-1241
Construct: endogenous protein (Jurkat)
Average holdup BI: 0.08
Immobilized partner concentration (10-6M): 10
P value: 1.06
Experiment method: nHU_Jurkat
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-08-03
Number of measurements: 3
The affinity was below the detection threshold of the assay.
PUBMED ID: 36542723
Experimental Details

-log10(P Two-sided unpaired T-test (6 vs 3))