Interaction Details


Details of experiments between Q9NZN5 and P27228
Note that the results of all experiments are listed, regardless of the modification states of the fragments.


Experiment series 1

Protein A

Protein: ARHGEF12
Fragment: 67-151
Site: PDZ
Construct: his6-MBP-TEVsite-PDZ

Protein B

Protein: HPV35-E6
Fragment: 140-149
Site: PBM
Sequence: SKPTRRETEV
Construct: biotin-ttds-SKPTRRETEV
Average holdup BI: 0.4
Immobilized partner concentration (10-6M): 39.7
Experiment method: CALIP_holdup
Details of affinity fitting: BSA and lysozyme internal standards + bacterial cell lysate + normal layout
Number of measurements: 1
Measured pKd: 4.24
PUBMED ID: 36115835

Experiment series 2

Protein A

Protein: ARHGEF12
Fragment: 67-151
Site: PDZ
Construct: his6-MBP-TEVsite-PDZ

Protein B

Protein: HPV35-E6
Fragment: 140-149
Site: PBM
Sequence: SKPTRRETEV
Construct: biotin-ttds-SKPTRRETEV
Average holdup BI: 0.15
Holdup BI standard deviation: 0.02
Normalized holdup BI: 0.17
Normalized holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 18.55
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: 4.06
PUBMED ID: 36115835

Experiment series 3

Protein A

Protein: ARHGEF12
Fragment: 67-151
Site: PDZ
Construct: his6-MBP-TEVsite-PDZ

Protein B

Protein: HPV35-E6
Fragment: 140-149
Site: PBM
Sequence: SKPTRRETEV
Construct: biotin-ttds-SKPTRRETEV
Measured dissociation constant (10-6M): 92.8
Standard deviation of dissociation constant (10-6M): 4.99
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit
Number of measurements: 3
Measured pKd: 4.03
PUBMED ID: 36115835

Experiment series 4

Protein A

Protein: ARHGEF12
Fragment: 67-151
Site: PDZ
Construct: his6-MBP-TEVsite-PDZ

Protein B

Protein: HPV35-E6
Fragment: 140-149
Site: PBM
Sequence: SKPTRRETEV
Construct: biotin-ttds-SKPTRRETEV
Average holdup BI: 0.09
Normalized holdup BI: 0.13
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: 3.91
PUBMED ID: 36115835
Experimental Details

not reported internal standards of holdup layout #6

Experiment series 5

Protein A

Protein: ARHGEF12
Fragment: 67-151
Site: PDZ
Construct: his6-MBP-TEVsite-PDZ

Protein B

Protein: HPV35-E6
Fragment: 140-149
Site: PBM
Sequence: SKPTRRETEV
Construct: biotin-ttds-SKPTRRETEV
Average holdup BI: 0.06
Normalized holdup BI: 0.08
Immobilized partner concentration (10-6M): 18.55
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: 3.68
Experimental Details

not reported internal standards of holdup layout #5

Experiment series 6

Protein A

Protein: ARHGEF12
Fragment: 67-151
Site: PDZ
Construct: his6-MBP-TEVsite-PDZ

Protein B

Protein: HPV35-E6
Fragment: 140-149
Site: PBM
Modification: (TPO)147
Sequence: SKPTRRETEV
Construct: Biotin-ttds-SKPTRREpTEV
Average holdup BI: -0.03
Normalized holdup BI: -0.04
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