Details of experiments between Q86UL8 and P50479
Note that the results of all experiments are listed, regardless of the modification states of the fragments.



Experiment series 1

Protein A protein: MAGI2
Protein A fragment: 772-865
Protein A site: PDZ_4
Protein A construct: his6-MBP-TEVsite-PDZ

Protein B protein: PDLIM4
Protein B fragment: 237-246
Protein B site: PBM
Protein B modification: iso2
Protein B sequence: VYPNAKVELV
Protein B construct: biotin-ado-ado-VYPNAKVELV

Average holdup BI: 0.04
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
Experimental details: Crude peptide synthesis product. The peptide concentration in the affinity conversion step may differ from reality causing a systematic error in reported affinities.
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 3.56


Experiment series 2

Protein A protein: MAGI2
Protein A fragment: 410-522
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ

Protein B protein: PDLIM4
Protein B fragment: 237-246
Protein B site: PBM
Protein B modification: iso2
Protein B sequence: VYPNAKVELV
Protein B construct: biotin-ado-ado-VYPNAKVELV

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 causing a systematic error in reported affinities.
PUBMED ID of publication: 36115835
Number of measurements: 1
The affinity was below the detection threshold of the assay.


Experiment series 3

Protein A protein: MAGI2
Protein A fragment: 600-682
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ

Protein B protein: PDLIM4
Protein B fragment: 237-246
Protein B site: PBM
Protein B modification: iso2
Protein B sequence: VYPNAKVELV
Protein B construct: biotin-ado-ado-VYPNAKVELV

Average holdup BI: 0.02
Normalized holdup BI: 0.03
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 causing a systematic error in reported affinities.
PUBMED ID of publication: 36115835
Number of measurements: 1
The affinity was below the detection threshold of the assay.


Experiment series 4

Protein A protein: MAGI2
Protein A fragment: 915-1017
Protein A site: PDZ_5
Protein A construct: his6-MBP-TEVsite-PDZ

Protein B protein: PDLIM4
Protein B fragment: 237-246
Protein B site: PBM
Protein B modification: iso2
Protein B sequence: VYPNAKVELV
Protein B construct: biotin-ado-ado-VYPNAKVELV

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 causing a systematic error in reported affinities.
PUBMED ID of publication: 36115835
Number of measurements: 1
The affinity was below the detection threshold of the assay.


Experiment series 5

Protein A protein: MAGI2
Protein A fragment: 1141-1229
Protein A site: PDZ_6
Protein A construct: his6-MBP-TEVsite-PDZ

Protein B protein: PDLIM4
Protein B fragment: 237-246
Protein B site: PBM
Protein B modification: iso2
Protein B sequence: VYPNAKVELV
Protein B construct: biotin-ado-ado-VYPNAKVELV

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 causing a systematic error in reported affinities.
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: MAGI2
Protein A fragment: 410-522
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ

Protein B protein: PDLIM4
Protein B fragment: 321-330
Protein B site: PBM
Protein B sequence: PSASCAVTAA
Protein B construct: Biotin-Ahx-PSASCAVTAA

Average holdup BI: -0.03
Holdup BI standard deviation: 0.01
Normalized holdup BI: -0.03
Normalized holdup BI standard deviation: 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
PUBMED ID of publication: 36115835
Number of measurements: 2
The affinity was below the detection threshold of the assay.


Experiment series 7

Protein A protein: MAGI2
Protein A fragment: 600-682
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ

Protein B protein: PDLIM4
Protein B fragment: 321-330
Protein B site: PBM
Protein B sequence: PSASCAVTAA
Protein B construct: Biotin-Ahx-PSASCAVTAA

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 8

Protein A protein: MAGI2
Protein A fragment: 772-865
Protein A site: PDZ_4
Protein A construct: his6-MBP-TEVsite-PDZ

Protein B protein: PDLIM4
Protein B fragment: 321-330
Protein B site: PBM
Protein B sequence: PSASCAVTAA
Protein B construct: Biotin-Ahx-PSASCAVTAA

Average holdup BI: -0.03
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 9

Protein A protein: MAGI2
Protein A fragment: 915-1017
Protein A site: PDZ_5
Protein A construct: his6-MBP-TEVsite-PDZ

Protein B protein: PDLIM4
Protein B fragment: 321-330
Protein B site: PBM
Protein B sequence: PSASCAVTAA
Protein B construct: Biotin-Ahx-PSASCAVTAA

Average holdup BI: -0.03
Normalized holdup BI: -0.03
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 10

Protein A protein: MAGI2
Protein A fragment: 1141-1229
Protein A site: PDZ_6
Protein A construct: his6-MBP-TEVsite-PDZ

Protein B protein: PDLIM4
Protein B fragment: 321-330
Protein B site: PBM
Protein B sequence: PSASCAVTAA
Protein B construct: Biotin-Ahx-PSASCAVTAA

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
PUBMED ID of publication: 36115835
Number of measurements: 1
The affinity was below the detection threshold of the assay.