Interaction Details


Details of experiments between Q14160 and Q96BN8
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: OTULIN
Fragment: 343-352
Site: PBM
Sequence: PVRVCEETSL
Construct: biotin-ttds-PVRVCEETSL
Experiment method: nHU-WB_Jurkat
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-08-03
Number of measurements: 3
Measured pKd: 6.22
PUBMED ID: 36542723

Experiment series 2

Protein A

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

Protein B

Protein: OTULIN
Fragment: 343-352
Site: PBM
Sequence: PVRVCEETSL
Construct: Biotin-ado-ado-PVRVCEETSL
Average holdup BI: 0.89
Normalized holdup BI: 0.89
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: 5.74
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: SCRIB
Fragment: 718-814
Site: PDZ_1
Construct: avi(biotin)-his6-MBP-TEVsite-PDZ

Protein B

Protein: OTULIN
Fragment: 1-352
Construct: endogenous protein (Jurkat)
Average holdup BI: 0.84
Immobilized partner concentration (10-6M): 10
P value: 4.74
Experiment method: nHU_Jurkat
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-08-03
Number of measurements: 3
Measured pKd: 5.73
PUBMED ID: 36542723
Experimental Details

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

Experiment series 4

Protein A

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

Protein B

Protein: OTULIN
Fragment: 343-352
Site: PBM
Sequence: PVRVCEETSL
Construct: biotin-ttds-PVRVCEETSL
Average holdup BI: 0.78
Normalized holdup BI: 0.86
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: 5.64
PUBMED ID: 36115835

Experiment series 5

Protein A

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

Protein B

Protein: OTULIN
Fragment: 343-352
Site: PBM
Sequence: PVRVCEETSL
Construct: biotin-ttds-PVRVCEETSL
Average holdup BI: 0.76
Normalized holdup BI: 0.8
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: 5.44
PUBMED ID: 36115835

Experiment series 6

Protein A

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

Protein B

Protein: OTULIN
Fragment: 343-352
Site: PBM
Sequence: PVRVCEETSL
Construct: Biotin-ado-ado-PVRVCEETSL
Average holdup BI: 0.64
Normalized holdup BI: 0.76
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: 5.32
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 7

Protein A

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

Protein B

Protein: OTULIN
Fragment: 343-352
Site: PBM
Sequence: PVRVCEETSL
Construct: biotin-ttds-PVRVCEETSL
Average holdup BI: 0.36
Holdup BI standard deviation: 0.02
Normalized holdup BI: 0.45
Normalized holdup BI standard deviation: 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: 2
Measured pKd: 4.7
PUBMED ID: 36115835

Experiment series 8

Protein A

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

Protein B

Protein: OTULIN
Fragment: 343-352
Site: PBM
Sequence: PVRVCEETSL
Construct: Biotin-ado-ado-PVRVCEETSL
Average holdup BI: 0.38
Normalized holdup BI: 0.38
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: 4.57
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: SCRIB
Fragment: 858-949
Site: PDZ_2
Construct: avi(biotin)-his6-MBP-TEVsite-PDZ

Protein B

Protein: OTULIN
Fragment: 1-352
Construct: endogenous protein (Jurkat)
Average holdup BI: 0.26
Immobilized partner concentration (10-6M): 10
P value: 4.41
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))