Interaction Details


Details of experiments between Q14160 and Q96KB5
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: his6-MBP-TEVsite-PDZ

Protein B

Protein: PBK
Fragment: 313-322
Site: PBM
Sequence: HIVEALETDV
Construct: biotin-ttds-HIVEALETDV
Average holdup BI: 0.46
Immobilized partner concentration (10-6M): 5.5
Experiment method: SAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normal layout
Number of measurements: 1
Measured pKd: 5.37
PUBMED ID: 36115835

Experiment series 2

Protein A

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

Protein B

Protein: PBK
Fragment: 313-322
Site: PBM
Sequence: HIVEALETDV
Construct: biotin-ttds-HIVEALETDV
Average holdup BI: 0.52
Normalized holdup BI: 0.65
Immobilized partner concentration (10-6M): 11.6
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: 5.32
PUBMED ID: 36115835

Experiment series 3

Protein A

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

Protein B

Protein: PBK
Fragment: 313-322
Site: PBM
Sequence: HIVEALETDV
Construct: biotin-ttds-HIVEALETDV
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 (10-6M): 11.6
Experiment method: DAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normalization based on CALIP_holdup data + reverse layout
Number of measurements: 4
Measured pKd: 5.32
PUBMED ID: 36115835

Experiment series 4

Protein A

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

Protein B

Protein: PBK
Fragment: 313-322
Site: PBM
Sequence: HIVEALETDV
Construct: biotin-ttds-HIVEALETDV
Average holdup BI: 0.39
Immobilized partner concentration (10-6M): 5.5
Experiment method: SAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normal layout
Number of measurements: 1
Measured pKd: 5.21
PUBMED ID: 36115835

Experiment series 5

Protein A

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

Protein B

Protein: PBK
Fragment: 313-322
Site: PBM
Sequence: HIVEALETDV
Construct: biotin-ttds-HIVEALETDV
Average holdup BI: 0.53
Normalized holdup BI: 0.59
Immobilized partner concentration (10-6M): 11.6
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: 5.18
PUBMED ID: 36115835

Experiment series 6

Protein A

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

Protein B

Protein: PBK
Fragment: 313-322
Site: PBM
Sequence: HIVEALETDV
Construct: biotin-ttds-HIVEALETDV
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
Number of measurements: 3
Measured pKd: 5.12
PUBMED ID: 36115835

Experiment series 7

Protein A

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

Protein B

Protein: PBK
Fragment: 313-322
Site: PBM
Sequence: HIVEALETDV
Construct: biotin-ttds-HIVEALETDV
Average holdup BI: 0.25
Immobilized partner concentration (10-6M): 5.5
Experiment method: SAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normal layout
Number of measurements: 1
Measured pKd: 4.86
PUBMED ID: 36115835

Experiment series 8

Protein A

Protein: SCRIB
Fragment: 1090-1197
Site: PDZ_4
Construct: his6-MBP-TEVsite-PDZ

Protein B

Protein: PBK
Fragment: 313-322
Site: PBM
Sequence: HIVEALETDV
Construct: biotin-ttds-HIVEALETDV
Average holdup BI: 0.07
Immobilized partner concentration (10-6M): 5.5
Experiment method: SAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normal layout
Number of measurements: 1
The affinity was below the detection threshold of the assay.
PUBMED ID: 36115835

Experiment series 9

Protein A

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

Protein B

Protein: PBK
Fragment: 1-322
Construct: endogenous protein (Jurkat)
Average holdup BI: -0.08
Immobilized partner concentration (10-6M): 10
P value: 0.38
Experiment method: nHU_Jurkat
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-08-03
Number of measurements: 3
The affinity was below the detection threshold of the assay.
PUBMED ID: 36542723
Experimental Details

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

Experiment series 10

Protein A

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

Protein B

Protein: PBK
Fragment: 1-322
Construct: endogenous protein (Jurkat)
Average holdup BI: -0.12
Immobilized partner concentration (10-6M): 10
P value: 0.69
Experiment method: nHU_Jurkat
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-08-03
Number of measurements: 3
The affinity was below the detection threshold of the assay.
PUBMED ID: 36542723
Experimental Details

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