Details of experiments between Q96BN8 and Q9C0E4
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: GRIP2
Protein B fragment: 934-1023
Protein B site: PDZ_7
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.08
Normalized holdup BI: 0.08
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: 3.66


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: GRIP2
Protein B fragment: 238-339
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.06
Normalized holdup BI: 0.06
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: 3.57


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: GRIP2
Protein B fragment: 446-548
Protein B site: PDZ_4
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.06
Normalized holdup BI: 0.06
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: 3.53


Experiment series 4

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: GRIP2
Protein B fragment: 649-740
Protein B site: PDZ_6
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.06
Normalized holdup BI: 0.06
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: 3.53


Experiment series 5

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: GRIP2
Protein B fragment: 544-642
Protein B site: PDZ_5
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.04
Normalized holdup BI: 0.04
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
The affinity was below the detection threshold of the assay.


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: GRIP2
Protein B fragment: 649-740
Protein B site: PDZ_6
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.02
Normalized holdup BI: 0.02
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
The affinity was below the detection threshold of the assay.


Experiment series 7

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: GRIP2
Protein B fragment: 544-642
Protein B site: PDZ_5
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.01
Normalized holdup BI: 0.01
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
The affinity was below the detection threshold of the assay.


Experiment series 8

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: GRIP2
Protein B fragment: 238-339
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: -0
Normalized holdup BI: -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
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
The affinity was below the detection threshold of the assay.


Experiment series 9

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: GRIP2
Protein B fragment: 934-1023
Protein B site: PDZ_7
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: -0.02
Normalized holdup BI: -0.02
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
The affinity was below the detection threshold of the assay.


Experiment series 10

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: GRIP2
Protein B fragment: 446-548
Protein B site: PDZ_4
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: -0.02
Normalized holdup BI: -0.02
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
The affinity was below the detection threshold of the assay.