Details of experiments between Q8TEW8 and Q15418
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: PARD3B
Protein A fragment: 184-299
Protein A site: PDZ_1
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: 0.33
Holdup BI standard deviation: 0.09
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: 4.1
Experiment series 2
Protein A protein: PARD3B
Protein A fragment: 376-471
Protein A site: PDZ_2
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: 0.03
Holdup BI standard deviation: 0.04
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: PARD3B
Protein A fragment: 490-594
Protein A site: PDZ_3
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: 0.08
Holdup BI standard deviation: 0.03
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 4
Protein A protein: PARD3B
Protein A fragment: 184-299
Protein A site: PDZ_1
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: 0.43
Holdup BI standard deviation: 0.14
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: 4.41
Experiment series 5
Protein A protein: PARD3B
Protein A fragment: 376-471
Protein A site: PDZ_2
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: -0.01
Holdup BI standard deviation: 0.05
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 6
Protein A protein: PARD3B
Protein A fragment: 490-594
Protein A site: PDZ_3
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: -0.03
Holdup BI standard deviation: 0.02
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 7
Protein A protein: PARD3B
Protein A fragment: 184-299
Protein A site: PDZ_1
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): 49.11
Standard deviation of dissociation constant (10-6M): 2.37
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 4.31
Experiment series 8
Protein A protein: PARD3B
Protein A fragment: 184-299
Protein A site: PDZ_1
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): 19.16
Standard deviation of dissociation constant (10-6M): 0.95
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 9
Protein A protein: PARD3B
Protein A fragment: 184-299
Protein A site: PDZ_1
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): 101.55
Standard deviation of dissociation constant (10-6M): 6.69
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 3.99
Experiment series 10
Protein A protein: PARD3B
Protein A fragment: 184-299
Protein A site: PDZ_1
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): 126.96
Standard deviation of dissociation constant (10-6M): 11.49
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 3.9
Experiment series 11
Protein A protein: PARD3B
Protein A fragment: 184-299
Protein A site: PDZ_1
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: 0.29
Holdup BI standard deviation: 0.03
Normalized holdup BI: 0.36
Normalized holdup BI standard deviation: 0.04
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: 4.62
Experiment series 12
Protein A protein: PARD3B
Protein A fragment: 184-299
Protein A site: PDZ_1
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: 0.1
Holdup BI standard deviation: 0
Normalized holdup BI: 0.12
Normalized holdup BI standard deviation: 0
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: 3.9
Experiment series 13
Protein A protein: PARD3B
Protein A fragment: 184-299
Protein A site: PDZ_1
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: 0.21
Normalized holdup BI: 0.21
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: 4.04
Experiment series 14
Protein A protein: PARD3B
Protein A fragment: 184-299
Protein A site: PDZ_1
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: 0.24
Holdup BI standard deviation: 0
Normalized holdup BI: 0.3
Normalized holdup BI standard deviation: 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: 2
Measured pKd: 4.4
Experiment series 15
Protein A protein: PARD3B
Protein A fragment: 184-299
Protein A site: PDZ_1
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: 0.4
Normalized holdup BI: 0.4
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: 4.69
Experiment series 16
Protein A protein: PARD3B
Protein A fragment: 184-299
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: RPS6KA1
Protein B fragment: 726-735
Protein B site: PBM
Protein B sequence: RVRKLPSTTL
Protein B construct: Biotin-ado-ado-RVRKLPSTTL
Average holdup BI: 0.45
Normalized holdup BI: 0.45
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: 4.7
Experiment series 17
Protein A protein: PARD3B
Protein A fragment: 184-299
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: RPS6KA1
Protein B fragment: 729-735
Protein B site: PBM
Protein B sequence: RVRKLPSTTL
Protein B construct: Fluorescein-KLPSTTL
Measured dissociation constant (10-6M): 27
Standard deviation of dissociation constant (10-6M): 3
Experiment method: FP_direct
PUBMED ID of publication: 30726710
Number of measurements: 3
Measured pKd: 4.57
Experiment series 18
Protein A protein: PARD3B
Protein A fragment: 184-299
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: RPS6KA1
Protein B fragment: 729-735
Protein B site: PBM
Protein B modification: (SEP)732
Protein B sequence: RVRKLPSTTL
Protein B construct: Fluorescein-KLP(pS)TTL
Measured dissociation constant (10-6M): 6.8
Standard deviation of dissociation constant (10-6M): 0.5
Experiment method: FP_direct
PUBMED ID of publication: 30726710
Number of measurements: 3
Measured pKd: 5.17
Experiment series 19
Protein A protein: PARD3B
Protein A fragment: 184-299
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: RPS6KA1
Protein B fragment: 683-735
Protein B site: PBM
Protein B sequence: RVRKLPSTTL
Protein B construct: ...KLPSTTL
Measured dissociation constant (10-6M): 45
Standard deviation of dissociation constant (10-6M): 7
Experiment method: FP_competitive
PUBMED ID of publication: 30726710
Number of measurements: 3
Measured pKd: 4.35
Experiment series 20
Protein A protein: PARD3B
Protein A fragment: 184-299
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: RPS6KA1
Protein B fragment: 683-735
Protein B site: PBM
Protein B modification: (SEP)732
Protein B sequence: RVRKLPSTTL
Protein B construct: ...KLP(pS)TTL
Measured dissociation constant (10-6M): 31
Standard deviation of dissociation constant (10-6M): 3.5
Experiment method: FP_competitive
PUBMED ID of publication: 30726710
Number of measurements: 3
Measured pKd: 4.51
Experiment series 21
Protein A protein: PARD3B
Protein A fragment: 184-299
Protein A site: PDZ_1
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): 4
Standard deviation of dissociation constant (10-6M): 0.3
Experiment method: SPR
PUBMED ID of publication: 30726710
Number of measurements: 1
Measured pKd: 5.4
Experiment series 22
Protein A protein: PARD3B
Protein A fragment: 184-299
Protein A site: PDZ_1
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): 6.1
Standard deviation of dissociation constant (10-6M): 0.4
Experiment method: SPR
PUBMED ID of publication: 30726710
Number of measurements: 1
Measured pKd: 5.21