Details of experiments between Q9BQQ3 and Q15418
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: GORASP1
Protein A fragment: 1-208
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: RPS6KA1
Protein B fragment: 725-735
Protein B site: PBM
Protein B modification: (SEP)732
Protein B sequence: RVRKLPSTTL
Protein B construct: biotin-ttds-RRVRKLPpSTTL
Average holdup BI: 0.03
Holdup BI standard deviation: 0.02
Immobilized partner concentration in holdup experiment (10-6M): 40
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: 3
The affinity was below the detection threshold of the assay.
Experiment series 2
Protein A protein: GORASP1
Protein A fragment: 1-208
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: RPS6KA1
Protein B fragment: 725-735
Protein B site: PBM
Protein B sequence: RVRKLPSTTL
Protein B construct: biotin-ttds-RRVRKLPSTTL
Average holdup BI: 0.01
Holdup BI standard deviation: 0.02
Immobilized partner concentration in holdup experiment (10-6M): 31
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: 4
The affinity was below the detection threshold of the assay.
Experiment series 3
Protein A protein: GORASP1
Protein A fragment: 1-208
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: RPS6KA1
Protein B fragment: 725-735
Protein B site: PBM
Protein B modification: (SEP)732
Protein B sequence: RVRKLPSTTL
Protein B construct: biotin-ttds-RRVRKLPpSTTL
Average holdup BI: 0
Normalized holdup BI: 0
Immobilized partner concentration in holdup experiment (10-6M): 15
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: GORASP1
Protein A fragment: 1-208
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: RPS6KA1
Protein B fragment: 725-735
Protein B site: PBM
Protein B modification: (TPO)733
Protein B sequence: RVRKLPSTTL
Protein B construct: biotin-ttds-RRVRKLPSpTTL
Average holdup BI: 0.08
Normalized holdup BI: 0.09
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
Measured pKd: 3.75
Experiment series 5
Protein A protein: GORASP1
Protein A fragment: 1-208
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: RPS6KA1
Protein B fragment: 725-735
Protein B site: PBM
Protein B modification: (TPO)734
Protein B sequence: RVRKLPSTTL
Protein B construct: Biotin-ttds-RRVRKLPSTpTL
Average holdup BI: 0.07
Normalized holdup BI: 0.09
Immobilized partner concentration in holdup experiment (10-6M): 25
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: 3.6
Experiment series 6
Protein A protein: GORASP1
Protein A fragment: 1-208
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: RPS6KA1
Protein B fragment: 725-735
Protein B site: PBM
Protein B sequence: RVRKLPSTTL
Protein B construct: biotin-ttds-RRVRKLPSTTL
Average holdup BI: 0.04
Normalized holdup BI: 0.05
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 7
Protein A protein: GORASP1
Protein A fragment: 1-208
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: RPS6KA1
Protein B fragment: 725-735
Protein B site: PBM
Protein B modification: (SEP)732
Protein B sequence: RVRKLPSTTL
Protein B construct: biotin-ttds-RRVRKLPpSTTL
Average holdup BI: 0.07
Normalized holdup BI: 0.08
Immobilized partner concentration in holdup experiment (10-6M): 15
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: 3.79
Experiment series 8
Protein A protein: GORASP1
Protein A fragment: 1-208
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: RPS6KA1
Protein B fragment: 726-735
Protein B site: PBM
Protein B sequence: RVRKLPSTTL
Protein B construct: Biotin-ado-ado-RVRKLPSTTL
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
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
The affinity was below the detection threshold of the assay.