Details of experiments between P03126 and Q5T5U3
Note that the results of all experiments are listed, regardless of the modification states of the fragments.



Experiment series 1

Protein A protein: HPV16-E6
Protein A fragment: 149-158
Protein A site: PBM
Protein A modification: (TPO)156
Protein A sequence: SSRTRRETQL
Protein A construct: Biotin-ttds-SSRTRREpTQL

Protein B protein: ARHGAP21
Protein B fragment: 47-159
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.2
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: CALIP_holdup
Details of affinity fitting: BSA and lysozyme internal standards + bacterial cell lysate + normal layout
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 4.17


Experiment series 2

Protein A protein: HPV16-E6
Protein A fragment: 149-158
Protein A site: PBM
Protein A sequence: SSRTRRETQL
Protein A construct: biotin-ttds-SSRTRRETQL

Protein B protein: ARHGAP21
Protein B fragment: 47-159
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0
Holdup BI standard deviation: 0.08
Immobilized partner concentration in holdup experiment (10-6M): 15
Experiment method: CALIP_holdup
Details of affinity fitting: BSA and lysozyme internal standards + bacterial cell lysate + normal layout
PUBMED ID of publication: 36115835
Number of measurements: 6
The affinity was below the detection threshold of the assay.


Experiment series 3

Protein A protein: HPV16-E6
Protein A fragment: 149-158
Protein A site: PBM
Protein A modification: (TPO)156
Protein A sequence: SSRTRRETQL
Protein A construct: Biotin-ttds-SSRTRREpTQL

Protein B protein: ARHGAP21
Protein B fragment: 47-159
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.16
Normalized holdup BI: 0.16
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: 4.05


Experiment series 4

Protein A protein: HPV16-E6
Protein A fragment: 149-158
Protein A site: PBM
Protein A sequence: SSRTRRETQL
Protein A construct: biotin-ttds-SSRTRRETQL

Protein B protein: ARHGAP21
Protein B fragment: 47-159
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.03
Holdup BI standard deviation: 0
Normalized holdup BI: 0.03
Normalized holdup BI standard deviation: 0
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: 2
The affinity was below the detection threshold of the assay.


Experiment series 5

Protein A protein: HPV16-E6
Protein A fragment: 149-158
Protein A site: PBM
Protein A sequence: SSRTRRETQL
Protein A construct: biotin-ttds-SSRTRRETQL

Protein B protein: ARHGAP21
Protein B fragment: 47-159
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.05
Normalized holdup BI: 0.05
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
Experimental details: not reported internal standards of holdup layout #5
Number of measurements: 1
The affinity was below the detection threshold of the assay.


Experiment series 6

Protein A protein: HPV16-E6
Protein A fragment: 149-158
Protein A site: PBM
Protein A sequence: SSRTRRETQL
Protein A construct: Biotin-ado-ado-SSRTRRETQL

Protein B protein: ARHGAP21
Protein B fragment: 47-159
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.13
Normalized holdup BI: 0.13
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
Experimental details: Crude peptide synthesis product. The peptide concentration in the affinity conversion step may differ from reality. Not reported internal standards of holdup layout #6
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 3.92