Details of experiments between Q9P202 and Q5T2W1
Note that the results of all experiments are listed, regardless of the modification states of the fragments.



Experiment series 1

Protein A protein: DFNB31
Protein A fragment: 565-578
Protein A site: Internal_PBM
Protein A construct: Biotin-GSGSGS-VSVDDVRSTSQGLSSY-CONH2

Protein B protein: NHERF3
Protein B fragment: 2-107
Protein B site: PDZ_1
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
The affinity was below the detection threshold of the assay.


Experiment series 2

Protein A protein: DFNB31
Protein A fragment: 565-578
Protein A site: Internal_PBM
Protein A construct: Biotin-GSGSGS-VSVDDVRSTSQGLSSY-CONH2

Protein B protein: NHERF3
Protein B fragment: 238-329
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: -0.1
Normalized holdup BI: -0.1
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 3

Protein A protein: DFNB31
Protein A fragment: 565-578
Protein A site: Internal_PBM
Protein A construct: Biotin-GSGSGS-VSVDDVRSTSQGLSSY-CONH2

Protein B protein: NHERF3
Protein B fragment: 375-459
Protein B site: PDZ_4
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
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: DFNB31
Protein A fragment: 898-907
Protein A site: PBM
Protein A sequence: FLVTEFNVML
Protein A construct: biotin-ado-ado-FLVTEFNVML

Protein B protein: NHERF3
Protein B fragment: 2-107
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.52
Normalized holdup BI: 0.54
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: 4.88


Experiment series 5

Protein A protein: DFNB31
Protein A fragment: 898-907
Protein A site: PBM
Protein A sequence: FLVTEFNVML
Protein A construct: biotin-ado-ado-FLVTEFNVML

Protein B protein: NHERF3
Protein B fragment: 375-459
Protein B site: PDZ_4
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.37
Normalized holdup BI: 0.37
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: 4.55


Experiment series 6

Protein A protein: DFNB31
Protein A fragment: 898-907
Protein A site: PBM
Protein A sequence: FLVTEFNVML
Protein A construct: biotin-ado-ado-FLVTEFNVML

Protein B protein: NHERF3
Protein B fragment: 238-329
Protein B site: PDZ_3
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
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 7

Protein A protein: DFNB31
Protein A fragment: 136-226
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ

Protein B protein: NHERF3
Protein B fragment: 510-519
Protein B site: PBM
Protein B sequence: SSSNSEDTEM
Protein B construct: Biotin-Ahx-SSSNSEDTEM

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.


Experiment series 8

Protein A protein: DFNB31
Protein A fragment: 263-379
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ

Protein B protein: NHERF3
Protein B fragment: 510-519
Protein B site: PBM
Protein B sequence: SSSNSEDTEM
Protein B construct: Biotin-Ahx-SSSNSEDTEM

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
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: DFNB31
Protein A fragment: 813-904
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ

Protein B protein: NHERF3
Protein B fragment: 510-519
Protein B site: PBM
Protein B sequence: SSSNSEDTEM
Protein B construct: Biotin-Ahx-SSSNSEDTEM

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