Details of experiments between Q15418 and Q6ZMN7
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: PDZRN4
Protein B fragment: 214-318
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.45
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: 5
Measured pKd: 4.33
Experiment series 2
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: PDZRN4
Protein B fragment: 389-498
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.09
Holdup BI standard deviation: 0.08
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 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: PDZRN4
Protein B fragment: 214-318
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.38
Holdup BI standard deviation: 0.11
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: 4.32
Experiment series 4
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: PDZRN4
Protein B fragment: 389-498
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.04
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 5
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: PDZRN4
Protein B fragment: 214-318
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Measured dissociation constant (10-6M): 63.7
Standard deviation of dissociation constant (10-6M): 3.4
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 4.2
Experiment series 6
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: PDZRN4
Protein B fragment: 214-318
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Measured dissociation constant (10-6M): 80.01
Standard deviation of dissociation constant (10-6M): 6.81
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 4.1
Experiment series 7
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: PDZRN4
Protein B fragment: 214-318
Protein B site: PDZ_1
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 8
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: PDZRN4
Protein B fragment: 214-318
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Measured dissociation constant (10-6M): 48.11
Standard deviation of dissociation constant (10-6M): 8.31
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 4.32
Experiment series 9
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: PDZRN4
Protein B fragment: 214-318
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.12
Holdup BI standard deviation: 0
Normalized holdup BI: 0.12
Normalized holdup BI standard deviation: 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: 2
Measured pKd: 3.98
Experiment series 10
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: PDZRN4
Protein B fragment: 214-318
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.01
Holdup BI standard deviation: 0.02
Normalized holdup BI: 0.01
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 11
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: PDZRN4
Protein B fragment: 214-318
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.27
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: 4.31
Experiment series 12
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: PDZRN4
Protein B fragment: 214-318
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.19
Holdup BI standard deviation: 0.02
Normalized holdup BI: 0.19
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: 4.13
Experiment series 13
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: PDZRN4
Protein B fragment: 214-318
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.15
Normalized holdup BI: 0.19
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: 4.2
Experiment series 14
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: PDZRN4
Protein B fragment: 214-318
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.23
Normalized holdup BI: 0.26
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: 4.31
Experiment series 15
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: PDZRN4
Protein B fragment: 214-318
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Measured dissociation constant (10-6M): 33
Standard deviation of dissociation constant (10-6M): 5
Experiment method: FP_direct
PUBMED ID of publication: 30726710
Number of measurements: 3
Measured pKd: 4.48
Experiment series 16
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: PDZRN4
Protein B fragment: 214-318
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Measured dissociation constant (10-6M): 14
Standard deviation of dissociation constant (10-6M): 2
Experiment method: FP_direct
PUBMED ID of publication: 30726710
Number of measurements: 3
Measured pKd: 4.85
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: PDZRN4
Protein B fragment: 214-318
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Measured dissociation constant (10-6M): 0.97
Standard deviation of dissociation constant (10-6M): 0.18
Experiment method: SPR
PUBMED ID of publication: 30726710
Number of measurements: 1
Measured pKd: 6.01
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: PDZRN4
Protein B fragment: 214-318
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Measured dissociation constant (10-6M): 7.1
Standard deviation of dissociation constant (10-6M): 0.9
Experiment method: SPR
PUBMED ID of publication: 30726710
Number of measurements: 1
Measured pKd: 5.15