Details of experiments between Q14CM0 and Q5TCQ9
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: MAGI3
Protein B fragment: 398-503
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.69
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
Measured pKd: 5.76


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: MAGI3
Protein B fragment: 1-114
Protein B site: PDZ_1
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: MAGI3
Protein B fragment: 572-655
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.06
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: MAGI3
Protein B fragment: 716-815
Protein B site: PDZ_4
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.02
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 5

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: MAGI3
Protein B fragment: 847-941
Protein B site: PDZ_5
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 6

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: MAGI3
Protein B fragment: 1014-1103
Protein B site: PDZ_6
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.04
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 7

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: MAGI3
Protein B fragment: 398-503
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.7
Holdup BI standard deviation: 0
Normalized holdup BI: 0.77
Normalized holdup BI standard deviation: 0
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: 2
Measured pKd: 5.27


Experiment series 8

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: MAGI3
Protein B fragment: 398-503
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.83
Normalized holdup BI: 0.88
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: 5.68