Interaction Details


Details of experiments between Q14160 and Q8TBC3
Note that the results of all experiments are listed, regardless of the modification states of the fragments.


Experiment series 1

Protein A

Protein: SCRIB
Fragment: 718-814
Site: PDZ_1
Construct: avi(biotin)-his6-MBP-TEVsite-PDZ

Protein B

Protein: SHKBP1
Fragment: 1-707
Construct: endogenous protein (Jurkat)
Average holdup BI: 0.9
Immobilized partner concentration (10-6M): 10
P value: 5.29
Experiment method: nHU_Jurkat
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-08-03
Number of measurements: 3
Measured pKd: 5.97
PUBMED ID: 36542723
Experimental Details

-log10(P Two-sided unpaired T-test (6 vs 3))

Experiment series 2

Protein A

Protein: SCRIB
Fragment: 858-949
Site: PDZ_2
Construct: avi(biotin)-his6-MBP-TEVsite-PDZ

Protein B

Protein: SHKBP1
Fragment: 1-707
Construct: endogenous protein (Jurkat)
Average holdup BI: 0.84
Immobilized partner concentration (10-6M): 10
P value: 5.8
Experiment method: nHU_Jurkat
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-08-03
Number of measurements: 3
Measured pKd: 5.72
PUBMED ID: 36542723
Experimental Details

-log10(P Two-sided unpaired T-test (6 vs 3))

Experiment series 3

Protein A

Protein: SCRIB
Fragment: 999-1093
Site: PDZ_3
Construct: his6-MBP-TEVsite-PDZ

Protein B

Protein: SHKBP1
Fragment: 698-707
Site: PBM
Sequence: PKMKLNETSF
Construct: Biotin-Ahx-PKMKLNETSF
Average holdup BI: 0.28
Normalized holdup BI: 0.34
Immobilized partner concentration (10-6M): 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 pKd: 4.49
PUBMED ID: 36115835
Experimental Details

The peptide stock solution had solubility issues. The peptide concentration in the affinity conversion step may differ from reality causing a systematic error in reported affinities.

Experiment series 4

Protein A

Protein: SCRIB
Fragment: 718-814
Site: PDZ_1
Construct: his6-MBP-TEVsite-PDZ

Protein B

Protein: SHKBP1
Fragment: 698-707
Site: PBM
Sequence: PKMKLNETSF
Construct: Biotin-Ahx-PKMKLNETSF
Average holdup BI: 0.21
Holdup BI standard deviation: 0.01
Normalized holdup BI: 0.26
Normalized holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 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 pKd: 4.31
PUBMED ID: 36115835
Experimental Details

The peptide stock solution had solubility issues. The peptide concentration in the affinity conversion step may differ from reality causing a systematic error in reported affinities.

Experiment series 5

Protein A

Protein: SCRIB
Fragment: 858-949
Site: PDZ_2
Construct: his6-MBP-TEVsite-PDZ

Protein B

Protein: SHKBP1
Fragment: 698-707
Site: PBM
Sequence: PKMKLNETSF
Construct: Biotin-Ahx-PKMKLNETSF
Average holdup BI: 0.02
Normalized holdup BI: 0.03
Immobilized partner concentration (10-6M): 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.
PUBMED ID: 36115835
Experimental Details

The peptide stock solution had solubility issues. The peptide concentration in the affinity conversion step may differ from reality causing a systematic error in reported affinities.