Details of experiments between Q14CM0 and O95049
Note that the results of all experiments are listed, regardless of the modification states of the fragments.



Experiment series 1

Protein A protein: FRMPD4
Protein A fragment: 1313-1322
Protein A site: PBM
Protein A sequence: RLPKIKETTV
Protein A construct: biotin-ttds-RLPKIKETTV

Protein B protein: TJP3
Protein B fragment: 1-109
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0
Immobilized partner concentration in holdup experiment (10-6M): 6.6
Experiment method: SAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normal 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: FRMPD4
Protein A fragment: 1313-1322
Protein A site: PBM
Protein A sequence: RLPKIKETTV
Protein A construct: biotin-ttds-RLPKIKETTV

Protein B protein: TJP3
Protein B fragment: 189-274
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.01
Immobilized partner concentration in holdup experiment (10-6M): 6.6
Experiment method: SAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normal 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: FRMPD4
Protein A fragment: 1313-1322
Protein A site: PBM
Protein A sequence: RLPKIKETTV
Protein A construct: biotin-ttds-RLPKIKETTV

Protein B protein: TJP3
Protein B fragment: 373-482
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0
Immobilized partner concentration in holdup experiment (10-6M): 6.6
Experiment method: SAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normal 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: FRMPD4
Protein A fragment: 1313-1322
Protein A site: PBM
Protein A sequence: RLPKIKETTV
Protein A construct: biotin-ttds-RLPKIKETTV

Protein B protein: TJP3
Protein B fragment: 373-482
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: -0.07
Normalized holdup BI: -0.07
Immobilized partner concentration in holdup experiment (10-6M): 21.6
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 5

Protein A protein: FRMPD4
Protein A fragment: 1313-1322
Protein A site: PBM
Protein A sequence: RLPKIKETTV
Protein A construct: Biotin-ado-ado-RLPKIKETTV

Protein B protein: TJP3
Protein B fragment: 373-482
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
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: 3.58


Experiment series 6

Protein A protein: FRMPD4
Protein A fragment: 67-160
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ

Protein B protein: TJP3
Protein B fragment: 910-919
Protein B site: PBM
Protein B sequence: GYDWGPATDL
Protein B construct: Biotin-Ahx-GYDWGPATDL

Average holdup BI: 0.09
Holdup BI standard deviation: 0
Normalized holdup BI: 0.11
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: 3.83