Details of experiments between the fragments Q13884~107-196 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: SNTB1
Protein A fragment: 107-196
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: 0.48
Holdup BI standard deviation: 0.07
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: 4.38
Experiment series 2
Protein A protein: SNTB1
Protein A fragment: 107-196
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: 0.2
Holdup BI standard deviation: 0.06
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: 3.92
Experiment series 3
Protein A protein: SNTB1
Protein A fragment: 107-196
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
Measured dissociation constant (10-6M): 55.5
Standard deviation of dissociation constant (10-6M): 20.02
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 4.26
Experiment series 4
Protein A protein: SNTB1
Protein A fragment: 107-196
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
Measured dissociation constant (10-6M): 19.19
Standard deviation of dissociation constant (10-6M): 1.39
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 4.72
Experiment series 5
Protein A protein: SNTB1
Protein A fragment: 107-196
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
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: SNTB1
Protein A fragment: 107-196
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
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: SNTB1
Protein A fragment: 107-196
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: 0.21
Holdup BI standard deviation: 0.09
Normalized holdup BI: 0.26
Normalized holdup BI standard deviation: 0.11
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: 4.39
Experiment series 8
Protein A protein: SNTB1
Protein A fragment: 107-196
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: 0.03
Holdup BI standard deviation: 0.08
Normalized holdup BI: 0.04
Normalized holdup BI standard deviation: 0.1
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: SNTB1
Protein A fragment: 107-196
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: 0.15
Normalized holdup BI: 0.17
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: 3.91
Experiment series 10
Protein A protein: SNTB1
Protein A fragment: 107-196
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: 0.16
Holdup BI standard deviation: 0.04
Normalized holdup BI: 0.19
Normalized holdup BI standard deviation: 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: 2
Measured pKd: 4.14
Experiment series 11
Protein A protein: SNTB1
Protein A fragment: 107-196
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: 0.3
Normalized holdup BI: 0.33
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: 4.56
Experiment series 12
Protein A protein: SNTB1
Protein A fragment: 107-196
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
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 13
Protein A protein: SNTB1
Protein A fragment: 107-196
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-RRVRKLP(pS)TTL
Measured dissociation constant (10-6M): 48
Standard deviation of dissociation constant (10-6M): 5
Experiment method: SPR
PUBMED ID of publication: 30726710
Number of measurements: 1
Measured pKd: 4.32