Details of experiments between P60484 and O15085
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: PTEN
Protein A fragment: 393-403
Protein A site: PBM
Protein A modification: (ALY)402
Protein A sequence: EDQHTQITKV
Protein A construct: biotin-ttds-DEDQHTQITKacV
Protein B protein: ARHGEF11
Protein B fragment: 41-126
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.43
Immobilized partner concentration in holdup experiment (10-6M): 29
Experiment method: CALIP_holdup
Details of affinity fitting: BSA and lysozyme internal standards + bacterial cell lysate + normal layout
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 4.44
Experiment series 2
Protein A protein: PTEN
Protein A fragment: 393-403
Protein A site: PBM
Protein A sequence: EDQHTQITKV
Protein A construct: biotin-ttds-DEDQHTQITKV
Protein B protein: ARHGEF11
Protein B fragment: 41-126
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 36
Experiment method: CALIP_holdup
Details of affinity fitting: BSA and lysozyme internal standards + bacterial cell lysate + 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: PTEN
Protein A fragment: 393-403
Protein A site: PBM
Protein A modification: (ALY)402
Protein A sequence: EDQHTQITKV
Protein A construct: biotin-ttds-DEDQHTQITKacV
Protein B protein: ARHGEF11
Protein B fragment: 41-126
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.54
Normalized holdup BI: 0.67
Immobilized partner concentration in holdup experiment (10-6M): 17
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
Measured pKd: 5.16
Experiment series 4
Protein A protein: PTEN
Protein A fragment: 393-403
Protein A site: PBM
Protein A sequence: EDQHTQITKV
Protein A construct: biotin-ttds-DEDQHTQITKV
Protein B protein: ARHGEF11
Protein B fragment: 41-126
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.22
Normalized holdup BI: 0.28
Immobilized partner concentration in holdup experiment (10-6M): 19
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
Measured pKd: 4.33
Experiment series 5
Protein A protein: PTEN
Protein A fragment: 393-403
Protein A site: PBM
Protein A modification: (ALY)402
Protein A sequence: EDQHTQITKV
Protein A construct: biotin-ttds-DEDQHTQITKacV
Protein B protein: ARHGEF11
Protein B fragment: 41-126
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.33
Normalized holdup BI: 0.44
Immobilized partner concentration in holdup experiment (10-6M): 17
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
Experimental details: not reported internal standards of holdup layout #5
Number of measurements: 1
Measured pKd: 4.7
Experiment series 6
Protein A protein: PTEN
Protein A fragment: 393-403
Protein A site: PBM
Protein A modification: (ALY)402
Protein A sequence: EDQHTQITKV
Protein A construct: biotin-ttds-DEDQHTQITKacV
Protein B protein: ARHGEF11
Protein B fragment: 41-126
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.34
Normalized holdup BI: 0.56
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: not reported internal standards of holdup layout #6
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 4.91
Experiment series 7
Protein A protein: PTEN
Protein A fragment: 394-403
Protein A site: PBM
Protein A sequence: EDQHTQITKV
Protein A construct: Biotin-ado-ado-EDQHTQITKV
Protein B protein: ARHGEF11
Protein B fragment: 41-126
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.16
Normalized holdup BI: 0.27
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: 4.35
Experiment series 8
Protein A protein: PTEN
Protein A fragment: 391-403
Protein A site: PBM
Protein A sequence: EDQHTQITKV
Protein A construct: Biotin-ttds-PFDEDQHTQITKV
Protein B protein: ARHGEF11
Protein B fragment: 41-126
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.11
Immobilized partner concentration in holdup experiment (10-6M): 17
Experiment method: CALIP_holdup
Details of affinity fitting: lysozyme internal standard
PUBMED ID of publication: 33382810
Number of measurements: 1
Measured pKd: 3.89