Details of experiments between Q96BN8 and P78352
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: OTULIN
Protein A fragment: 343-352
Protein A site: PBM
Protein A sequence: PVRVCEETSL
Protein A construct: biotin-ttds-PVRVCEETSL
Protein B protein: DLG4
Protein B fragment: 155-249
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.17
Normalized holdup BI: 0.21
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 4.2
Experiment series 2
Protein A protein: OTULIN
Protein A fragment: 343-352
Protein A site: PBM
Protein A sequence: PVRVCEETSL
Protein A construct: biotin-ttds-PVRVCEETSL
Protein B protein: DLG4
Protein B fragment: 62-152
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.15
Normalized holdup BI: 0.18
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 4.11
Experiment series 3
Protein A protein: OTULIN
Protein A fragment: 343-352
Protein A site: PBM
Protein A sequence: PVRVCEETSL
Protein A construct: biotin-ttds-PVRVCEETSL
Protein B protein: DLG4
Protein B fragment: 306-403
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.12
Holdup BI standard deviation: 0
Normalized holdup BI: 0.14
Normalized holdup BI standard deviation: 0
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
PUBMED ID of publication: 36115835
Number of measurements: 2
Measured pKd: 3.96
Experiment series 4
Protein A protein: OTULIN
Protein A fragment: 343-352
Protein A site: PBM
Protein A sequence: PVRVCEETSL
Protein A construct: Biotin-ado-ado-PVRVCEETSL
Protein B protein: DLG4
Protein B fragment: 306-403
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.06
Normalized holdup BI: 0.07
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
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
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 3.62
Experiment series 5
Protein A protein: OTULIN
Protein A fragment: 343-352
Protein A site: PBM
Protein A sequence: PVRVCEETSL
Protein A construct: Biotin-ado-ado-PVRVCEETSL
Protein B protein: DLG4
Protein B fragment: 155-249
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.14
Normalized holdup BI: 0.17
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
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
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 4.08
Experiment series 6
Protein A protein: OTULIN
Protein A fragment: 343-352
Protein A site: PBM
Protein A sequence: PVRVCEETSL
Protein A construct: Biotin-ado-ado-PVRVCEETSL
Protein B protein: DLG4
Protein B fragment: 62-152
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.19
Normalized holdup BI: 0.23
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
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
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 4.24