Interaction Details


Details of experiments between Q96BN8 and Q96L92
Note that the results of all experiments are listed, regardless of the modification states of the fragments.


Experiment series 1

Protein A

Protein: OTULIN
Fragment: 1-352
Construct: endogenous protein (Jurkat)

Protein B

Protein: SNX27
Fragment: 40-141
Site: PDZ
Construct: avi(biotin)-his6-MBP-TEVsite-PDZ
Average holdup BI: 0.94
Immobilized partner concentration (10-6M): 10
P value: 4.76
Experiment method: nHU_Jurkat
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-08-03
Number of measurements: 3
Measured pKd: 6.2
PUBMED ID: 36542723
Experimental Details

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

Experiment series 2

Protein A

Protein: OTULIN
Fragment: 343-352
Site: PBM
Sequence: PVRVCEETSL
Construct: biotin-ttds-PVRVCEETSL

Protein B

Protein: SNX27
Fragment: 1-541
Construct: endogenous protein (Jurkat)
Experiment method: nHU-WB_Jurkat
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-08-03
Number of measurements: 3
Measured pKd: 5.83
PUBMED ID: 36542723

Experiment series 3

Protein A

Protein: OTULIN
Fragment: 1-352
Construct: endogenous protein (Jurkat)

Protein B

Protein: SNX27
Fragment: 40-141
Site: PDZ
Construct: avi(biotin)-his6-MBP-TEVsite-PDZ
Average holdup BI: 0.86
Immobilized partner concentration (10-6M): 10
P value: 4.92
Experiment method: nHU_Jurkat
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-06-27
Number of measurements: 6
Measured pKd: 5.78
PUBMED ID: 36542723
Experimental Details

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

Experiment series 4

Protein A

Protein: OTULIN
Fragment: 343-352
Site: PBM
Sequence: PVRVCEETSL
Construct: Biotin-ado-ado-PVRVCEETSL

Protein B

Protein: SNX27
Fragment: 40-141
Site: PDZ
Construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.88
Normalized holdup BI: 0.88
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: 5.72
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 5

Protein A

Protein: OTULIN
Fragment: 343-352
Site: PBM
Sequence: PVRVCEETSL
Construct: biotin-ttds-PVRVCEETSL

Protein B

Protein: SNX27
Fragment: 40-141
Site: PDZ
Construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.85
Normalized holdup BI: 0.85
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: 5.58
PUBMED ID: 36115835

Experiment series 6

Protein A

Protein: OTULIN
Fragment: 1-352
Construct: endogenous protein (Jurkat)

Protein B

Protein: SNX27
Fragment: 1-541
Construct: avi(biotin)-his6-MBP-TEVsite-SNX27
Average holdup BI: 0.03
Immobilized partner concentration (10-6M): 10
P value: 0.04
Experiment method: nHU_Jurkat
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-06-27
Number of measurements: 6
The affinity was below the detection threshold of the assay.
PUBMED ID: 36542723
Experimental Details

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