Details of experiments between Q15418 and O14907
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: RPS6KA1
Protein A fragment: 725-735
Protein A site: PBM
Protein A modification: (SEP)732
Protein A sequence: RVRKLPSTTL
Protein A construct: biotin-ttds-RRVRKLPpSTTL
Protein B protein: TX1BP3
Protein B fragment: 7-124
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.11
Holdup BI standard deviation: 0.05
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
Measured pKd: 3.47
Experiment series 2
Protein A protein: RPS6KA1
Protein A fragment: 725-735
Protein A site: PBM
Protein A sequence: RVRKLPSTTL
Protein A construct: biotin-ttds-RRVRKLPSTTL
Protein B protein: TX1BP3
Protein B fragment: 7-124
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.03
Holdup BI standard deviation: 0.04
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: RPS6KA1
Protein A fragment: 725-735
Protein A site: PBM
Protein A modification: (SEP)732
Protein A sequence: RVRKLPSTTL
Protein A construct: biotin-ttds-RRVRKLPpSTTL
Protein B protein: TX1BP3
Protein B fragment: 7-124
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.03
Holdup BI standard deviation: 0.04
Normalized holdup BI: 0.03
Normalized holdup BI standard deviation: 0.05
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: 2
The affinity was below the detection threshold of the assay.
Experiment series 4
Protein A protein: RPS6KA1
Protein A fragment: 725-735
Protein A site: PBM
Protein A modification: (TPO)733
Protein A sequence: RVRKLPSTTL
Protein A construct: biotin-ttds-RRVRKLPSpTTL
Protein B protein: TX1BP3
Protein B fragment: 7-124
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: -0.02
Holdup BI standard deviation: 0.04
Normalized holdup BI: -0.03
Normalized holdup BI standard deviation: 0.04
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: 2
The affinity was below the detection threshold of the assay.
Experiment series 5
Protein A protein: RPS6KA1
Protein A fragment: 725-735
Protein A site: PBM
Protein A modification: (TPO)734
Protein A sequence: RVRKLPSTTL
Protein A construct: Biotin-ttds-RRVRKLPSTpTL
Protein B protein: TX1BP3
Protein B fragment: 7-124
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.14
Normalized holdup BI: 0.15
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: 3.87
Experiment series 6
Protein A protein: RPS6KA1
Protein A fragment: 725-735
Protein A site: PBM
Protein A sequence: RVRKLPSTTL
Protein A construct: biotin-ttds-RRVRKLPSTTL
Protein B protein: TX1BP3
Protein B fragment: 7-124
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: -0.04
Holdup BI standard deviation: 0.08
Normalized holdup BI: -0.04
Normalized holdup BI standard deviation: 0.08
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: 2
The affinity was below the detection threshold of the assay.
Experiment series 7
Protein A protein: RPS6KA1
Protein A fragment: 725-735
Protein A site: PBM
Protein A modification: (SEP)732
Protein A sequence: RVRKLPSTTL
Protein A construct: biotin-ttds-RRVRKLPpSTTL
Protein B protein: TX1BP3
Protein B fragment: 7-124
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.05
Normalized holdup BI: 0.06
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: 3.62
Experiment series 8
Protein A protein: RPS6KA1
Protein A fragment: 726-735
Protein A site: PBM
Protein A sequence: RVRKLPSTTL
Protein A construct: Biotin-ado-ado-RVRKLPSTTL
Protein B protein: TX1BP3
Protein B fragment: 7-124
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ
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.
Experiment series 9
Protein A protein: RPS6KA1
Protein A fragment: 1-735
Protein A construct: endogenous protein (Jurkat)
Protein B protein: TX1BP3
Protein B fragment: 7-124
Protein B site: PDZ
Protein B construct: avi(biotin)-his6-MBP-TEVsite-PDZ
Average holdup BI: -0.05
Immobilized partner concentration in holdup experiment (10-6M): 10
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 1.18
Experiment method: nHU_Jurkat
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.08.03.
Experimental details: -log10(P Two-sided unpaired T-test (6 vs 3))
PUBMED ID of publication: 36542723
Number of measurements: 3
The affinity was below the detection threshold of the assay.