Interaction Details


Details of experiments between Q14160 and Q13574
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: DGKZ
Fragment: 919-928
Site: PBM
Sequence: IQREDQETAV
Construct: biotin-ttds-IQREDQETAV
Experiment method: nHU-WB_Jurkat
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-08-03
Number of measurements: 3
Measured pKd: 6.42
PUBMED ID: 36542723

Experiment series 2

Protein A

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

Protein B

Protein: DGKZ
Fragment: 919-928
Site: PBM
Sequence: IQREDQETAV
Construct: biotin-ttds-IQREDQETAV
Experiment method: nHU-WB_Jurkat
Details of affinity fitting: Hill equation (8 point titration) h=0.6
Date of measurement: 2022-08-03
Number of measurements: 3
Measured pKd: 6.4
PUBMED ID: 36542723

Experiment series 3

Protein A

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

Protein B

Protein: DGKZ
Fragment: 919-928
Site: PBM
Sequence: IQREDQETAV
Construct: biotin-ttds-IQREDQETAV
Average holdup BI: 0.5
Holdup BI standard deviation: 0.03
Normalized holdup BI: 0.56
Normalized holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 13.5
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: 5.05
PUBMED ID: 36115835

Experiment series 4

Protein A

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

Protein B

Protein: DGKZ
Fragment: 919-928
Site: PBM
Sequence: IQREDQETAV
Construct: biotin-ttds-IQREDQETAV
Average holdup BI: 0.47
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 5

Protein A

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

Protein B

Protein: DGKZ
Fragment: 919-928
Site: PBM
Sequence: IQREDQETAV
Construct: biotin-ttds-IQREDQETAV
Average holdup BI: 0.37
Holdup BI standard deviation: 0.02
Normalized holdup BI: 0.46
Normalized holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 13.5
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
Number of measurements: 3
Measured pKd: 4.86
PUBMED ID: 36115835

Experiment series 6

Protein A

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

Protein B

Protein: DGKZ
Fragment: 919-928
Site: PBM
Sequence: IQREDQETAV
Construct: biotin-ttds-IQREDQETAV
Average holdup BI: 0.25
Immobilized partner concentration (10-6M): 6
Experiment method: SAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normal layout
Number of measurements: 1
Measured pKd: 4.81
PUBMED ID: 36115835

Experiment series 7

Protein A

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

Protein B

Protein: DGKZ
Fragment: 919-928
Site: PBM
Sequence: IQREDQETAV
Construct: biotin-ttds-IQREDQETAV
Average holdup BI: 0.36
Holdup BI standard deviation: 0.24
Normalized holdup BI: 0.38
Normalized holdup BI standard deviation: 0.26
Immobilized partner concentration (10-6M): 13.5
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
Number of measurements: 5
Measured pKd: 4.71
PUBMED ID: 36115835

Experiment series 8

Protein A

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

Protein B

Protein: DGKZ
Fragment: 919-928
Site: PBM
Sequence: IQREDQETAV
Construct: biotin-ttds-IQREDQETAV
Average holdup BI: 0.45
Normalized holdup BI: 0.45
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: 4.71
PUBMED ID: 36115835
Experimental Details

not reported internal standards of holdup layout #6

Experiment series 9

Protein A

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

Protein B

Protein: DGKZ
Fragment: 919-928
Site: PBM
Sequence: IQREDQETAV
Construct: biotin-ttds-IQREDQETAV
Average holdup BI: 0.32
Normalized holdup BI: 0.32
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: 4.45
PUBMED ID: 36115835
Experimental Details

not reported internal standards of holdup layout #6

Experiment series 10

Protein A

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

Protein B

Protein: DGKZ
Fragment: 919-928
Site: PBM
Sequence: IQREDQETAV
Construct: biotin-ttds-IQREDQETAV
Measured dissociation constant (10-6M): 36.87
Standard deviation of dissociation constant (10-6M): 3.4
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
Number of measurements: 3
Measured pKd: 4.43
PUBMED ID: 36115835

Experiment series 11

Protein A

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

Protein B

Protein: DGKZ
Fragment: 919-928
Site: PBM
Sequence: IQREDQETAV
Construct: biotin-ttds-IQREDQETAV
Average holdup BI: 0.12
Immobilized partner concentration (10-6M): 6
Experiment method: SAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normal layout
Number of measurements: 1
Measured pKd: 4.4
PUBMED ID: 36115835

Experiment series 12

Protein A

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

Protein B

Protein: DGKZ
Fragment: 919-928
Site: PBM
Sequence: IQREDQETAV
Construct: biotin-ttds-IQREDQETAV
Average holdup BI: 0.11
Immobilized partner concentration (10-6M): 6
Experiment method: SAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normal layout
Number of measurements: 1
Measured pKd: 4.36
PUBMED ID: 36115835

Experiment series 13

Protein A

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

Protein B

Protein: DGKZ
Fragment: 919-928
Site: PBM
Sequence: IQREDQETAV
Construct: biotin-ttds-IQREDQETAV
Average holdup BI: 0.3
Immobilized partner concentration (10-6M): 10
P value: 3.71
Experiment method: nHU_Jurkat
Details of affinity fitting: hyperbolic binding equation
Number of measurements: 3
The affinity was below the detection threshold of the assay.
PUBMED ID: 37942667
Experimental Details

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

Experiment series 14

Protein A

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

Protein B

Protein: DGKZ
Fragment: 1-928
Construct: endogenous protein (Jurkat)
Average holdup BI: -0.09
Immobilized partner concentration (10-6M): 10
P value: 0.43
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 15

Protein A

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

Protein B

Protein: DGKZ
Fragment: 1-928
Construct: endogenous protein (Jurkat)
Average holdup BI: -0.06
Immobilized partner concentration (10-6M): 10
P value: 0.27
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 16

Protein A

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

Protein B

Protein: DGKZ
Fragment: 919-928
Site: PBM
Sequence: IQREDQETAV
Construct: biotin-ttds-IQREDQETAV
Average holdup BI: 0.01
Immobilized partner concentration (10-6M): 6
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