Details of experiments between Q5T2W1 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: NHERF3 Fragment: 2-107 Site: PDZ_1 Construct: his6-MBP-TEVsite-PDZ
Protein B Protein: RPS6KA1 Fragment: 725-735 Site: PBM Sequence: RVRKLPSTTL Construct: biotin-ttds-RRVRKLPSTTL
Average holdup BI: 0.42
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6 M): 31
Experiment method: CALIP_holdup
Details of affinity fitting: BSA and lysozyme internal standards + bacterial cell lysate + normal layout
Number of measurements: 4
Measured pK d : 4.4
Experiment series 2 Protein A Protein: NHERF3 Fragment: 2-107 Site: PDZ_1 Construct: his6-MBP-TEVsite-PDZ
Protein B Protein: RPS6KA1 Fragment: 726-735 Site: PBM Sequence: RVRKLPSTTL Construct: Biotin-ado-ado-RVRKLPSTTL
Average holdup BI: 0.18
Normalized holdup BI: 0.19
Immobilized partner concentration (10-6 M): 18
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
Number of measurements: 1
Measured pK d : 4.14
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
Experiment series 3 Protein A Protein: NHERF3 Fragment: 2-107 Site: PDZ_1 Construct: his6-MBP-TEVsite-PDZ
Protein B Protein: RPS6KA1 Fragment: 729-735 Site: PBM Sequence: RVRKLPSTTL Construct: Fluorescein-KLPSTTL
Measured dissociation constant (10-6 M): 80
Standard deviation of dissociation constant (10-6 M): 20
Experiment method: FP_direct
Number of measurements: 3
Measured pK d : 4.1
Experiment series 4 Protein A Protein: NHERF3 Fragment: 2-107 Site: PDZ_1 Construct: his6-MBP-TEVsite-PDZ
Protein B Protein: RPS6KA1 Fragment: 725-735 Site: PBM Sequence: RVRKLPSTTL Construct: biotin-ttds-RRVRKLPSTTL
Average holdup BI: 0.15
Holdup BI standard deviation: 0.01
Normalized holdup BI: 0.16
Normalized holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6 M): 18
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
Number of measurements: 2
Measured pK d : 4.03
Experiment series 5 Protein A Protein: NHERF3 Fragment: 375-459 Site: PDZ_4 Construct: his6-MBP-TEVsite-PDZ
Protein B Protein: RPS6KA1 Fragment: 725-735 Site: PBM Sequence: RVRKLPSTTL Construct: biotin-ttds-RRVRKLPSTTL
Average holdup BI: 0.07
Normalized holdup BI: 0.07
Immobilized partner concentration (10-6 M): 18
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
Number of measurements: 1
Measured pK d : 3.65
Experiment series 6 Protein A Protein: NHERF3 Fragment: 238-329 Site: PDZ_3 Construct: his6-MBP-TEVsite-PDZ
Protein B Protein: RPS6KA1 Fragment: 725-735 Site: PBM Sequence: RVRKLPSTTL Construct: biotin-ttds-RRVRKLPSTTL
Average holdup BI: 0.07
Normalized holdup BI: 0.07
Immobilized partner concentration (10-6 M): 18
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
Number of measurements: 1
Measured pK d : 3.61
Experiment series 7 Protein A Protein: NHERF3 Fragment: 132-224 Site: PDZ_2 Construct: his6-MBP-TEVsite-PDZ
Protein B Protein: RPS6KA1 Fragment: 725-735 Site: PBM Sequence: RVRKLPSTTL Construct: biotin-ttds-RRVRKLPSTTL
Average holdup BI: -0.03
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6 M): 31
Experiment method: CALIP_holdup
Details of affinity fitting: BSA and lysozyme internal standards + bacterial cell lysate + normal layout
Number of measurements: 4
The affinity was below the detection threshold of the assay.
Experiment series 8 Protein A Protein: NHERF3 Fragment: 2-107 Site: PDZ_1 Construct: his6-MBP-TEVsite-PDZ
Protein B Protein: RPS6KA1 Fragment: 725-735 Site: PBM Sequence: RVRKLPSTTL Construct: Biotin-ttds-RRVRKLPSTTL
Measured dissociation constant (10-6 M): 0
Experiment method: SPR
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 9 Protein A Protein: NHERF3 Fragment: 2-107 Site: PDZ_1 Construct: his6-MBP-TEVsite-PDZ
Protein B Protein: RPS6KA1 Fragment: 725-735 Site: PBM Sequence: RVRKLPSTTL Construct: biotin-ttds-RRVRKLPSTTL
Measured dissociation constant (10-6 M): 0
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
Number of measurements: 3
The affinity was below the detection threshold of the assay.
Experiment series 10 Protein A Protein: NHERF3 Fragment: 238-329 Site: PDZ_3 Construct: his6-MBP-TEVsite-PDZ
Protein B Protein: RPS6KA1 Fragment: 725-735 Site: PBM Sequence: RVRKLPSTTL Construct: biotin-ttds-RRVRKLPSTTL
Average holdup BI: 0
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6 M): 31
Experiment method: CALIP_holdup
Details of affinity fitting: BSA and lysozyme internal standards + bacterial cell lysate + normal layout
Number of measurements: 4
The affinity was below the detection threshold of the assay.
Experiment series 11 Protein A Protein: NHERF3 Fragment: 238-329 Site: PDZ_3 Construct: his6-MBP-TEVsite-PDZ
Protein B Protein: RPS6KA1 Fragment: 726-735 Site: PBM Sequence: RVRKLPSTTL Construct: Biotin-ado-ado-RVRKLPSTTL
Average holdup BI: -0.01
Normalized holdup BI: -0.01
Immobilized partner concentration (10-6 M): 18
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
Number of measurements: 1
The affinity was below the detection threshold of the assay.
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
Experiment series 12 Protein A Protein: NHERF3 Fragment: 375-459 Site: PDZ_4 Construct: his6-MBP-TEVsite-PDZ
Protein B Protein: RPS6KA1 Fragment: 725-735 Site: PBM Sequence: RVRKLPSTTL Construct: biotin-ttds-RRVRKLPSTTL
Average holdup BI: -0.03
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6 M): 31
Experiment method: CALIP_holdup
Details of affinity fitting: BSA and lysozyme internal standards + bacterial cell lysate + normal layout
Number of measurements: 4
The affinity was below the detection threshold of the assay.
Experiment series 13 Protein A Protein: NHERF3 Fragment: 375-459 Site: PDZ_4 Construct: his6-MBP-TEVsite-PDZ
Protein B Protein: RPS6KA1 Fragment: 726-735 Site: PBM Sequence: RVRKLPSTTL Construct: Biotin-ado-ado-RVRKLPSTTL
Average holdup BI: 0.02
Normalized holdup BI: 0.02
Immobilized partner concentration (10-6 M): 18
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
Number of measurements: 1
The affinity was below the detection threshold of the assay.
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
Experiment series 14 Protein A Protein: NHERF3 Fragment: 2-107 Site: PDZ_1 Construct: his6-MBP-TEVsite-PDZ
Protein B Protein: RPS6KA1 Fragment: 725-735 Site: PBM Modification: (SEP)732 Sequence: RVRKLPSTTL Construct: biotin-ttds-RRVRKLPpSTTL
Average holdup BI: 0.1
Normalized holdup BI: 0.11
Immobilized partner concentration (10-6 M): 15
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
Number of measurements: 1
Measured pK d : 3.92
Experimental Details not reported internal standards of holdup layout #5
Experiment series 15 Protein A Protein: NHERF3 Fragment: 375-459 Site: PDZ_4 Construct: his6-MBP-TEVsite-PDZ
Protein B Protein: RPS6KA1 Fragment: 725-735 Site: PBM Modification: (SEP)732 Sequence: RVRKLPSTTL Construct: biotin-ttds-RRVRKLPpSTTL
Average holdup BI: 0.07
Normalized holdup BI: 0.07
Immobilized partner concentration (10-6 M): 15
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
Number of measurements: 1
Measured pK d : 3.7
Experimental Details not reported internal standards of holdup layout #5
Experiment series 16 Protein A Protein: NHERF3 Fragment: 2-107 Site: PDZ_1 Construct: his6-MBP-TEVsite-PDZ
Protein B Protein: RPS6KA1 Fragment: 729-735 Site: PBM Modification: (SEP)732 Sequence: RVRKLPSTTL Construct: Fluorescein-KLP(pS)TTL
Measured dissociation constant (10-6 M): 220
Standard deviation of dissociation constant (10-6 M): 30
Experiment method: FP_direct
Number of measurements: 3
Measured pK d : 3.66
Experiment series 17 Protein A Protein: NHERF3 Fragment: 2-107 Site: PDZ_1 Construct: his6-MBP-TEVsite-PDZ
Protein B Protein: RPS6KA1 Fragment: 725-735 Site: PBM Modification: (SEP)732 Sequence: RVRKLPSTTL Construct: biotin-ttds-RRVRKLPpSTTL
Average holdup BI: 0.11
Holdup BI standard deviation: 0.14
Immobilized partner concentration (10-6 M): 40
Experiment method: CALIP_holdup
Details of affinity fitting: BSA and lysozyme internal standards + bacterial cell lysate + normal layout
Number of measurements: 3
Measured pK d : 3.49
Experiment series 18 Protein A Protein: NHERF3 Fragment: 132-224 Site: PDZ_2 Construct: his6-MBP-TEVsite-PDZ
Protein B Protein: RPS6KA1 Fragment: 725-735 Site: PBM Modification: (SEP)732 Sequence: RVRKLPSTTL Construct: biotin-ttds-RRVRKLPpSTTL
Average holdup BI: 0.01
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6 M): 40
Experiment method: CALIP_holdup
Details of affinity fitting: BSA and lysozyme internal standards + bacterial cell lysate + normal layout
Number of measurements: 3
The affinity was below the detection threshold of the assay.
Experiment series 19 Protein A Protein: NHERF3 Fragment: 2-107 Site: PDZ_1 Construct: his6-MBP-TEVsite-PDZ
Protein B Protein: RPS6KA1 Fragment: 725-735 Site: PBM Modification: (SEP)732 Sequence: RVRKLPSTTL Construct: Biotin-ttds-RRVRKLP(pS)TTL
Measured dissociation constant (10-6 M): 0
Experiment method: SPR
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 20 Protein A Protein: NHERF3 Fragment: 2-107 Site: PDZ_1 Construct: his6-MBP-TEVsite-PDZ
Protein B Protein: RPS6KA1 Fragment: 725-735 Site: PBM Modification: (SEP)732 Sequence: RVRKLPSTTL Construct: biotin-ttds-RRVRKLPpSTTL
Average holdup BI: -0.01
Holdup BI standard deviation: 0.01
Normalized holdup BI: -0.01
Normalized holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6 M): 15
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
Number of measurements: 2
The affinity was below the detection threshold of the assay.
Experiment series 21 Protein A Protein: NHERF3 Fragment: 2-107 Site: PDZ_1 Construct: his6-MBP-TEVsite-PDZ
Protein B Protein: RPS6KA1 Fragment: 725-735 Site: PBM Modification: (SEP)732 Sequence: RVRKLPSTTL Construct: biotin-ttds-RRVRKLPpSTTL
Measured dissociation constant (10-6 M): 0
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
Number of measurements: 3
The affinity was below the detection threshold of the assay.
Experiment series 22 Protein A Protein: NHERF3 Fragment: 238-329 Site: PDZ_3 Construct: his6-MBP-TEVsite-PDZ
Protein B Protein: RPS6KA1 Fragment: 725-735 Site: PBM Modification: (SEP)732 Sequence: RVRKLPSTTL Construct: biotin-ttds-RRVRKLPpSTTL
Average holdup BI: 0.02
Normalized holdup BI: 0.02
Immobilized partner concentration (10-6 M): 15
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 23 Protein A Protein: NHERF3 Fragment: 238-329 Site: PDZ_3 Construct: his6-MBP-TEVsite-PDZ
Protein B Protein: RPS6KA1 Fragment: 725-735 Site: PBM Modification: (SEP)732 Sequence: RVRKLPSTTL Construct: biotin-ttds-RRVRKLPpSTTL
Average holdup BI: 0.04
Normalized holdup BI: 0.04
Immobilized partner concentration (10-6 M): 15
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experimental Details not reported internal standards of holdup layout #5
Experiment series 24 Protein A Protein: NHERF3 Fragment: 238-329 Site: PDZ_3 Construct: his6-MBP-TEVsite-PDZ
Protein B Protein: RPS6KA1 Fragment: 725-735 Site: PBM Modification: (SEP)732 Sequence: RVRKLPSTTL Construct: biotin-ttds-RRVRKLPpSTTL
Average holdup BI: 0.04
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6 M): 40
Experiment method: CALIP_holdup
Details of affinity fitting: BSA and lysozyme internal standards + bacterial cell lysate + normal layout
Number of measurements: 3
The affinity was below the detection threshold of the assay.
Experiment series 25 Protein A Protein: NHERF3 Fragment: 375-459 Site: PDZ_4 Construct: his6-MBP-TEVsite-PDZ
Protein B Protein: RPS6KA1 Fragment: 725-735 Site: PBM Modification: (SEP)732 Sequence: RVRKLPSTTL Construct: biotin-ttds-RRVRKLPpSTTL
Average holdup BI: 0.01
Normalized holdup BI: 0.01
Immobilized partner concentration (10-6 M): 15
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 26 Protein A Protein: NHERF3 Fragment: 375-459 Site: PDZ_4 Construct: his6-MBP-TEVsite-PDZ
Protein B Protein: RPS6KA1 Fragment: 725-735 Site: PBM Modification: (SEP)732 Sequence: RVRKLPSTTL Construct: biotin-ttds-RRVRKLPpSTTL
Average holdup BI: 0.03
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6 M): 40
Experiment method: CALIP_holdup
Details of affinity fitting: BSA and lysozyme internal standards + bacterial cell lysate + normal layout
Number of measurements: 3
The affinity was below the detection threshold of the assay.
Experiment series 27 Protein A Protein: NHERF3 Fragment: 2-107 Site: PDZ_1 Construct: his6-MBP-TEVsite-PDZ
Protein B Protein: RPS6KA1 Fragment: 725-735 Site: PBM Modification: (TPO)733 Sequence: RVRKLPSTTL Construct: biotin-ttds-RRVRKLPSpTTL
Average holdup BI: -0
Holdup BI standard deviation: 0
Normalized holdup BI: -0
Normalized holdup BI standard deviation: 0
Immobilized partner concentration (10-6 M): 18
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
Number of measurements: 2
The affinity was below the detection threshold of the assay.
Experiment series 28 Protein A Protein: NHERF3 Fragment: 2-107 Site: PDZ_1 Construct: his6-MBP-TEVsite-PDZ
Protein B Protein: RPS6KA1 Fragment: 725-735 Site: PBM Modification: (TPO)733 Sequence: RVRKLPSTTL Construct: biotin-ttds-RRVRKLPSpTTL
Measured dissociation constant (10-6 M): 0
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
Number of measurements: 3
The affinity was below the detection threshold of the assay.
Experiment series 29 Protein A Protein: NHERF3 Fragment: 238-329 Site: PDZ_3 Construct: his6-MBP-TEVsite-PDZ
Protein B Protein: RPS6KA1 Fragment: 725-735 Site: PBM Modification: (TPO)733 Sequence: RVRKLPSTTL Construct: biotin-ttds-RRVRKLPSpTTL
Average holdup BI: 0.02
Normalized holdup BI: 0.02
Immobilized partner concentration (10-6 M): 18
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 30 Protein A Protein: NHERF3 Fragment: 375-459 Site: PDZ_4 Construct: his6-MBP-TEVsite-PDZ
Protein B Protein: RPS6KA1 Fragment: 725-735 Site: PBM Modification: (TPO)733 Sequence: RVRKLPSTTL Construct: biotin-ttds-RRVRKLPSpTTL
Average holdup BI: -0.01
Normalized holdup BI: -0.01
Immobilized partner concentration (10-6 M): 18
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 31 Protein A Protein: NHERF3 Fragment: 2-107 Site: PDZ_1 Construct: his6-MBP-TEVsite-PDZ
Protein B Protein: RPS6KA1 Fragment: 725-735 Site: PBM Modification: (TPO)734 Sequence: RVRKLPSTTL Construct: Biotin-ttds-RRVRKLPSTpTL
Average holdup BI: 0.25
Normalized holdup BI: 0.28
Immobilized partner concentration (10-6 M): 25
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
Number of measurements: 1
Measured pK d : 4.2
Experiment series 32 Protein A Protein: NHERF3 Fragment: 375-459 Site: PDZ_4 Construct: his6-MBP-TEVsite-PDZ
Protein B Protein: RPS6KA1 Fragment: 725-735 Site: PBM Modification: (TPO)734 Sequence: RVRKLPSTTL Construct: Biotin-ttds-RRVRKLPSTpTL
Average holdup BI: 0.12
Normalized holdup BI: 0.12
Immobilized partner concentration (10-6 M): 25
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
Number of measurements: 1
Measured pK d : 3.75
Experiment series 33 Protein A Protein: NHERF3 Fragment: 238-329 Site: PDZ_3 Construct: his6-MBP-TEVsite-PDZ
Protein B Protein: RPS6KA1 Fragment: 725-735 Site: PBM Modification: (TPO)734 Sequence: RVRKLPSTTL Construct: Biotin-ttds-RRVRKLPSTpTL
Average holdup BI: 0.05
Normalized holdup BI: 0.05
Immobilized partner concentration (10-6 M): 25
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
Number of measurements: 1
Measured pK d : 3.36
Experiment series 34 Protein A Protein: NHERF3 Fragment: 2-107 Site: PDZ_1 Construct: his6-MBP-TEVsite-PDZ
Protein B Protein: RPS6KA1 Fragment: 725-735 Site: PBM Modification: (TPO)734 Sequence: RVRKLPSTTL Construct: Biotin-ttds-RRVRKLPSTpTL
Measured dissociation constant (10-6 M): 0
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
Number of measurements: 3
The affinity was below the detection threshold of the assay.
Please cite ProfAff! (Gogl, G. et al., Nature Communications 2022)
© The ProfAff database is developped by the research team of Gergo Gogl .