Details of experiments between Q12959 and Q13613
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: DLG1
Protein A fragment: 314-412
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: MTMR1
Protein B fragment: 656-665
Protein B site: PBM
Protein B sequence: HSATSVHTSV
Protein B construct: Biotin-Ahx-HSATSVHTSV
Average holdup BI: 0.46
Normalized holdup BI: 0.56
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: 1
Measured pKd: 4.9
Experiment series 2
Protein A protein: DLG1
Protein A fragment: 221-314
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: MTMR1
Protein B fragment: 656-665
Protein B site: PBM
Protein B sequence: HSATSVHTSV
Protein B construct: Biotin-Ahx-HSATSVHTSV
Average holdup BI: 0.18
Normalized holdup BI: 0.24
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: 1
Measured pKd: 4.26
Experiment series 3
Protein A protein: DLG1
Protein A fragment: 457-570
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: MTMR1
Protein B fragment: 656-665
Protein B site: PBM
Protein B sequence: HSATSVHTSV
Protein B construct: Biotin-Ahx-HSATSVHTSV
Average holdup BI: 0.05
Normalized holdup BI: 0.06
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: 1
Measured pKd: 3.56
Experiment series 4
Protein A protein: DLG1
Protein A fragment: 221-314
Protein A site: PDZ_1
Protein A construct: avi(biotin)-his6-MBP-TEVsite-PDZ
Protein B protein: MTMR1
Protein B fragment: 1-665
Protein B construct: endogenous protein (Jurkat)
Average holdup BI: 0.24
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): 4.43
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 5
Protein A protein: DLG1
Protein A fragment: 314-412
Protein A site: PDZ_2
Protein A construct: avi(biotin)-his6-MBP-TEVsite-PDZ
Protein B protein: MTMR1
Protein B fragment: 1-665
Protein B construct: endogenous protein (Jurkat)
Average holdup BI: 0.9
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): 6.19
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
Measured pKd: 5.97