Details of experiments between the fragments O15085~41-126 and Q15418~725-735
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: ARHGEF11
Protein A fragment: 41-126
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: 0.13
Holdup BI standard deviation: 0.13
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: 4
Measured pKd: 3.57
Experiment series 2
Protein A protein: ARHGEF11
Protein A fragment: 41-126
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: 0.44
Holdup BI standard deviation: 0.13
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: 5
Measured pKd: 4.44
Experiment series 3
Protein A protein: ARHGEF11
Protein A fragment: 41-126
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: 0.02
Normalized holdup BI: 0.02
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: ARHGEF11
Protein A fragment: 41-126
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
Normalized holdup BI: -0.01
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 5
Protein A protein: ARHGEF11
Protein A fragment: 41-126
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: 0.24
Normalized holdup BI: 0.32
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: 4.29
Experiment series 6
Protein A protein: ARHGEF11
Protein A fragment: 41-126
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: 0.25
Normalized holdup BI: 0.32
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: 4.44
Experiment series 7
Protein A protein: ARHGEF11
Protein A fragment: 41-126
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: 0.06
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: 3.77