Details of experiments between Q15418 and Q13424
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: SNTA1
Protein B fragment: 84-171
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.47
Holdup BI standard deviation: 0.11
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: 4.36
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: SNTA1
Protein B fragment: 84-171
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.27
Holdup BI standard deviation: 0.09
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
Measured pKd: 4.09
Experiment series 3
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: SNTA1
Protein B fragment: 84-171
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ
Measured dissociation constant (10-6M): 182.67
Standard deviation of dissociation constant (10-6M): 17.71
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 3.74
Experiment series 4
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: SNTA1
Protein B fragment: 84-171
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ
Measured dissociation constant (10-6M): 63.95
Standard deviation of dissociation constant (10-6M): 4.71
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 4.19
Experiment series 5
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: SNTA1
Protein B fragment: 84-171
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ
Measured dissociation constant (10-6M): 0
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
PUBMED ID of publication: 36115835
Number of measurements: 3
The affinity was below the detection threshold of the assay.
Experiment series 6
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: SNTA1
Protein B fragment: 84-171
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ
Measured dissociation constant (10-6M): 0
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
PUBMED ID of publication: 36115835
Number of measurements: 3
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: SNTA1
Protein B fragment: 84-171
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.22
Holdup BI standard deviation: 0.03
Normalized holdup BI: 0.26
Normalized holdup BI standard deviation: 0.03
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
Measured pKd: 4.4
Experiment series 8
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: SNTA1
Protein B fragment: 84-171
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.03
Holdup BI standard deviation: 0.01
Normalized holdup BI: 0.04
Normalized holdup BI standard deviation: 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
PUBMED ID of publication: 36115835
Number of measurements: 2
The affinity was below the detection threshold of the assay.
Experiment series 9
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: SNTA1
Protein B fragment: 84-171
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.11
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: 3.85
Experiment series 10
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: SNTA1
Protein B fragment: 84-171
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.1
Holdup BI standard deviation: 0.01
Normalized holdup BI: 0.12
Normalized holdup BI standard deviation: 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
PUBMED ID of publication: 36115835
Number of measurements: 2
Measured pKd: 3.87
Experiment series 11
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: SNTA1
Protein B fragment: 84-171
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.22
Normalized holdup BI: 0.29
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: 4.47
Experiment series 12
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: SNTA1
Protein B fragment: 84-171
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.13
Normalized holdup BI: 0.15
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
Experiment series 13
Protein A protein: RPS6KA1
Protein A fragment: 729-735
Protein A site: PBM
Protein A sequence: RVRKLPSTTL
Protein A construct: Fluorescein-KLPSTTL
Protein B protein: SNTA1
Protein B fragment: 84-171
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ
Measured dissociation constant (10-6M): 41
Standard deviation of dissociation constant (10-6M): 11
Experiment method: FP_direct
PUBMED ID of publication: 30726710
Number of measurements: 3
Measured pKd: 4.39
Experiment series 14
Protein A protein: RPS6KA1
Protein A fragment: 729-735
Protein A site: PBM
Protein A modification: (SEP)732
Protein A sequence: RVRKLPSTTL
Protein A construct: Fluorescein-KLP(pS)TTL
Protein B protein: SNTA1
Protein B fragment: 84-171
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ
Measured dissociation constant (10-6M): 4.9
Standard deviation of dissociation constant (10-6M): 0.4
Experiment method: FP_direct
PUBMED ID of publication: 30726710
Number of measurements: 3
Measured pKd: 5.31
Experiment series 15
Protein A protein: RPS6KA1
Protein A fragment: 683-735
Protein A site: PBM
Protein A sequence: RVRKLPSTTL
Protein A construct: ...KLPSTTL
Protein B protein: SNTA1
Protein B fragment: 84-171
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ
Measured dissociation constant (10-6M): 81
Standard deviation of dissociation constant (10-6M): 17
Experiment method: FP_competitive
PUBMED ID of publication: 30726710
Number of measurements: 3
Measured pKd: 4.09
Experiment series 16
Protein A protein: RPS6KA1
Protein A fragment: 683-735
Protein A site: PBM
Protein A modification: (SEP)732
Protein A sequence: RVRKLPSTTL
Protein A construct: ...KLP(pS)TTL
Protein B protein: SNTA1
Protein B fragment: 84-171
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ
Measured dissociation constant (10-6M): 10.5
Standard deviation of dissociation constant (10-6M): 2.6
Experiment method: FP_competitive
PUBMED ID of publication: 30726710
Number of measurements: 3
Measured pKd: 4.98
Experiment series 17
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: SNTA1
Protein B fragment: 84-171
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ
Measured dissociation constant (10-6M): 0
Experiment method: SPR
PUBMED ID of publication: 30726710
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 18
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-RRVRKLP(pS)TTL
Protein B protein: SNTA1
Protein B fragment: 84-171
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ
Measured dissociation constant (10-6M): 90
Standard deviation of dissociation constant (10-6M): 4
Experiment method: SPR
PUBMED ID of publication: 30726710
Number of measurements: 1
Measured pKd: 4.05