Interaction Details


Details of experiments between Q14160 and Q15311
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: 999-1093
Site: PDZ_3
Construct: his6-MBP-TEVsite-PDZ

Protein B

Protein: RALBP1
Fragment: 646-655
Site: PBM
Sequence: PSRDRKETSI
Construct: biotin-ttds-PSRDRKETSI
Average holdup BI: 0.62
Normalized holdup BI: 0.73
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: 5.25
PUBMED ID: 36115835
Experimental Details

not reported internal standards of holdup layout #6

Experiment series 2

Protein A

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

Protein B

Protein: RALBP1
Fragment: 646-655
Site: PBM
Sequence: PSRDRKETSI
Construct: biotin-ttds-PSRDRKETSI
Average holdup BI: 0.63
Normalized holdup BI: 0.7
Immobilized partner concentration (10-6M): 28
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: 4.97
PUBMED ID: 36115835

Experiment series 3

Protein A

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

Protein B

Protein: RALBP1
Fragment: 646-655
Site: PBM
Sequence: PSRDRKETSI
Construct: biotin-ttds-PSRDRKETSI
Average holdup BI: 0.5
Normalized holdup BI: 0.5
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: 4.79
PUBMED ID: 36115835
Experimental Details

not reported internal standards of holdup layout #6

Experiment series 4

Protein A

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

Protein B

Protein: RALBP1
Fragment: 646-655
Site: PBM
Sequence: PSRDRKETSI
Construct: biotin-ttds-PSRDRKETSI
Measured dissociation constant (10-6M): 25.83
Standard deviation of dissociation constant (10-6M): 1.95
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
Number of measurements: 3
Measured pKd: 4.59
PUBMED ID: 36115835

Experiment series 5

Protein A

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

Protein B

Protein: RALBP1
Fragment: 646-655
Site: PBM
Sequence: PSRDRKETSI
Construct: biotin-ttds-PSRDRKETSI
Average holdup BI: 0.43
Holdup BI standard deviation: 0.02
Normalized holdup BI: 0.45
Normalized holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 28
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: 4.49
PUBMED ID: 36115835

Experiment series 6

Protein A

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

Protein B

Protein: RALBP1
Fragment: 646-655
Site: PBM
Sequence: PSRDRKETSI
Construct: biotin-ttds-PSRDRKETSI
Average holdup BI: 0.2
Immobilized partner concentration (10-6M): 20
Experiment method: SAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normal layout
Number of measurements: 1
Measured pKd: 4.12
PUBMED ID: 36115835

Experiment series 7

Protein A

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

Protein B

Protein: RALBP1
Fragment: 646-655
Site: PBM
Sequence: PSRDRKETSI
Construct: biotin-ttds-PSRDRKETSI
Average holdup BI: 0.12
Normalized holdup BI: 0.12
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: 3.91
PUBMED ID: 36115835
Experimental Details

not reported internal standards of holdup layout #6

Experiment series 8

Protein A

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

Protein B

Protein: RALBP1
Fragment: 646-655
Site: PBM
Sequence: PSRDRKETSI
Construct: biotin-ttds-PSRDRKETSI
Average holdup BI: 0.11
Normalized holdup BI: 0.13
Immobilized partner concentration (10-6M): 28
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: 3.75
PUBMED ID: 36115835

Experiment series 9

Protein A

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

Protein B

Protein: RALBP1
Fragment: 646-655
Site: PBM
Sequence: PSRDRKETSI
Construct: biotin-ttds-PSRDRKETSI
Average holdup BI: 0.03
Immobilized partner concentration (10-6M): 20
Experiment method: SAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normal layout
Number of measurements: 1
The affinity was below the detection threshold of the assay.
PUBMED ID: 36115835

Experiment series 10

Protein A

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

Protein B

Protein: RALBP1
Fragment: 1-655
Construct: endogenous protein (Jurkat)
Average holdup BI: 0.21
Immobilized partner concentration (10-6M): 10
P value: 1.66
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 11

Protein A

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

Protein B

Protein: RALBP1
Fragment: 1-655
Construct: endogenous protein (Jurkat)
Average holdup BI: 0.04
Immobilized partner concentration (10-6M): 10
P value: 0.23
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 12

Protein A

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

Protein B

Protein: RALBP1
Fragment: 646-655
Site: PBM
Sequence: PSRDRKETSI
Construct: biotin-ttds-PSRDRKETSI
Average holdup BI: 0.06
Immobilized partner concentration (10-6M): 20
Experiment method: SAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normal layout
Number of measurements: 1
The affinity was below the detection threshold of the assay.
PUBMED ID: 36115835

Experiment series 13

Protein A

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

Protein B

Protein: RALBP1
Fragment: 646-655
Site: PBM
Sequence: PSRDRKETSI
Construct: biotin-ttds-PSRDRKETSI
Average holdup BI: 0.08
Immobilized partner concentration (10-6M): 20
Experiment method: SAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normal layout
Number of measurements: 1
The affinity was below the detection threshold of the assay.
PUBMED ID: 36115835