Interaction Details


Details of experiments between Q6ZMN7 and Q14CM0
Note that the results of all experiments are listed, regardless of the modification states of the fragments.


Experiment series 1

Protein A

Protein: PDZRN4
Fragment: 214-318
Site: PDZ_1
Construct: his6-MBP-TEVsite-PDZ

Protein B

Protein: FRMPD4
Fragment: 1313-1322
Site: PBM
Sequence: RLPKIKETTV
Construct: Biotin-ado-ado-RLPKIKETTV
Average holdup BI: 0.53
Normalized holdup BI: 0.59
Immobilized partner concentration (10-6M): 18
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
Number of measurements: 1
Measured pKd: 4.96
PUBMED ID: 36115835
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

Experiment series 2

Protein A

Protein: PDZRN4
Fragment: 214-318
Site: PDZ_1
Construct: his6-MBP-TEVsite-PDZ

Protein B

Protein: FRMPD4
Fragment: 1313-1322
Site: PBM
Sequence: RLPKIKETTV
Construct: biotin-ttds-RLPKIKETTV
Measured dissociation constant (10-6M): 19.74
Standard deviation of dissociation constant (10-6M): 0.94
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
Number of measurements: 3
Measured pKd: 4.7
PUBMED ID: 36115835

Experiment series 3

Protein A

Protein: PDZRN4
Fragment: 214-318
Site: PDZ_1
Construct: his6-MBP-TEVsite-PDZ

Protein B

Protein: FRMPD4
Fragment: 1313-1322
Site: PBM
Sequence: RLPKIKETTV
Construct: biotin-ttds-RLPKIKETTV
Average holdup BI: 0.48
Holdup BI standard deviation: 0.03
Normalized holdup BI: 0.48
Normalized holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 21.6
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
Number of measurements: 3
Measured pKd: 4.67
PUBMED ID: 36115835

Experiment series 4

Protein A

Protein: PDZRN4
Fragment: 389-498
Site: PDZ_2
Construct: his6-MBP-TEVsite-PDZ

Protein B

Protein: FRMPD4
Fragment: 1313-1322
Site: PBM
Sequence: RLPKIKETTV
Construct: biotin-ttds-RLPKIKETTV
Average holdup BI: 0.14
Immobilized partner concentration (10-6M): 6.6
Experiment method: SAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normal layout
Number of measurements: 1
Measured pKd: 4.43
PUBMED ID: 36115835

Experiment series 5

Protein A

Protein: PDZRN4
Fragment: 1027-1036
Site: PBM
Sequence: HNAFLSVTTV
Construct: Biotin-Ahx-HNAFLSVTTV

Protein B

Protein: FRMPD4
Fragment: 67-160
Site: PDZ
Construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.05
Holdup BI standard deviation: 0.01
Normalized holdup BI: 0.05
Normalized holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 18
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
Number of measurements: 2
Measured pKd: 3.5
PUBMED ID: 36115835
Experimental Details

The peptide stock solution had solubility issues. The peptide concentration in the affinity conversion step may differ from reality causing a systematic error in reported affinities.

Experiment series 6

Protein A

Protein: PDZRN4
Fragment: 214-318
Site: PDZ_1
Construct: his6-MBP-TEVsite-PDZ

Protein B

Protein: FRMPD4
Fragment: 1313-1322
Site: PBM
Sequence: RLPKIKETTV
Construct: biotin-ttds-RLPKIKETTV
Average holdup BI: 0.07
Immobilized partner concentration (10-6M): 6.6
Experiment method: SAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normal layout
Number of measurements: 1
The affinity was below the detection threshold of the assay.
PUBMED ID: 36115835