Details of experiments between Q96KB5 and Q14160
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: PBK
Protein A fragment: 313-322
Protein A site: PBM
Protein A sequence: HIVEALETDV
Protein A construct: biotin-ttds-HIVEALETDV
Protein B protein: SCRIB
Protein B fragment: 718-814
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Measured dissociation constant (10-6M): 7.53
Standard deviation of dissociation constant (10-6M): 0.39
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 5.12
Experiment series 2
Protein A protein: PBK
Protein A fragment: 313-322
Protein A site: PBM
Protein A sequence: HIVEALETDV
Protein A construct: biotin-ttds-HIVEALETDV
Protein B protein: SCRIB
Protein B fragment: 718-814
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.46
Immobilized partner concentration in holdup experiment (10-6M): 5.5
Experiment method: SAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normal layout
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 5.37
Experiment series 3
Protein A protein: PBK
Protein A fragment: 313-322
Protein A site: PBM
Protein A sequence: HIVEALETDV
Protein A construct: biotin-ttds-HIVEALETDV
Protein B protein: SCRIB
Protein B fragment: 999-1093
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.39
Immobilized partner concentration in holdup experiment (10-6M): 5.5
Experiment method: SAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normal layout
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 5.21
Experiment series 4
Protein A protein: PBK
Protein A fragment: 313-322
Protein A site: PBM
Protein A sequence: HIVEALETDV
Protein A construct: biotin-ttds-HIVEALETDV
Protein B protein: SCRIB
Protein B fragment: 858-949
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.25
Immobilized partner concentration in holdup experiment (10-6M): 5.5
Experiment method: SAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normal layout
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 4.86
Experiment series 5
Protein A protein: PBK
Protein A fragment: 313-322
Protein A site: PBM
Protein A sequence: HIVEALETDV
Protein A construct: biotin-ttds-HIVEALETDV
Protein B protein: SCRIB
Protein B fragment: 1090-1197
Protein B site: PDZ_4
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.07
Immobilized partner concentration in holdup experiment (10-6M): 5.5
Experiment method: SAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normal layout
PUBMED ID of publication: 36115835
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 6
Protein A protein: PBK
Protein A fragment: 313-322
Protein A site: PBM
Protein A sequence: HIVEALETDV
Protein A construct: biotin-ttds-HIVEALETDV
Protein B protein: SCRIB
Protein B fragment: 858-949
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.52
Normalized holdup BI: 0.65
Immobilized partner concentration in holdup experiment (10-6M): 11.6
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: 5.32
Experiment series 7
Protein A protein: PBK
Protein A fragment: 313-322
Protein A site: PBM
Protein A sequence: HIVEALETDV
Protein A construct: biotin-ttds-HIVEALETDV
Protein B protein: SCRIB
Protein B fragment: 718-814
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.62
Holdup BI standard deviation: 0.03
Normalized holdup BI: 0.65
Normalized holdup BI standard deviation: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 11.6
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: 4
Measured pKd: 5.32
Experiment series 8
Protein A protein: PBK
Protein A fragment: 313-322
Protein A site: PBM
Protein A sequence: HIVEALETDV
Protein A construct: biotin-ttds-HIVEALETDV
Protein B protein: SCRIB
Protein B fragment: 999-1093
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.53
Normalized holdup BI: 0.59
Immobilized partner concentration in holdup experiment (10-6M): 11.6
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: 5.18
Experiment series 9
Protein A protein: PBK
Protein A fragment: 1-322
Protein A construct: endogenous protein (Jurkat)
Protein B protein: SCRIB
Protein B fragment: 718-814
Protein B site: PDZ_1
Protein B construct: avi(biotin)-his6-MBP-TEVsite-PDZ
Average holdup BI: -0.08
Immobilized partner concentration in holdup experiment (10-6M): 10
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.38
Experiment method: nHU_Jurkat
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.08.03.
Experimental details: -log10(P Two-sided unpaired T-test (6 vs 3))
PUBMED ID of publication: 36542723
Number of measurements: 3
The affinity was below the detection threshold of the assay.
Experiment series 10
Protein A protein: PBK
Protein A fragment: 1-322
Protein A construct: endogenous protein (Jurkat)
Protein B protein: SCRIB
Protein B fragment: 858-949
Protein B site: PDZ_2
Protein B construct: avi(biotin)-his6-MBP-TEVsite-PDZ
Average holdup BI: -0.12
Immobilized partner concentration in holdup experiment (10-6M): 10
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 0.69
Experiment method: nHU_Jurkat
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.08.03.
Experimental details: -log10(P Two-sided unpaired T-test (6 vs 3))
PUBMED ID of publication: 36542723
Number of measurements: 3
The affinity was below the detection threshold of the assay.