Details of experiments between P03126 and Q6ZMN7
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: PDZRN4
Protein B fragment: 389-498
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.24
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: 4.26


Experiment series 2

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: PDZRN4
Protein B fragment: 214-318
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.02
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
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 sequence: SSRTRRETQL
Protein A construct: biotin-ttds-SSRTRRETQL

Protein B protein: PDZRN4
Protein B fragment: 214-318
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.62
Holdup BI standard deviation: 0.16
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: 5
Measured pKd: 5.12


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: PDZRN4
Protein B fragment: 389-498
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.01
Holdup BI standard deviation: 0.11
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 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: PDZRN4
Protein B fragment: 214-318
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ

Measured dissociation constant (10-6M): 5.94
Standard deviation of dissociation constant (10-6M): 0.24
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit

PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 5.23


Experiment series 6

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: PDZRN4
Protein B fragment: 214-318
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ

Measured dissociation constant (10-6M): 0
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit

PUBMED ID of publication: 36115835
Number of measurements: 3
The affinity was below the detection threshold of the assay.


Experiment series 7

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: PDZRN4
Protein B fragment: 214-318
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: -0.09
Normalized holdup BI: -0.1
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
The affinity was below the detection threshold of the assay.


Experiment series 8

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: PDZRN4
Protein B fragment: 214-318
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.73
Holdup BI standard deviation: 0.02
Normalized holdup BI: 0.73
Normalized holdup BI standard deviation: 0.02
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
Measured pKd: 5.25


Experiment series 9

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: PDZRN4
Protein B fragment: 214-318
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.53
Normalized holdup BI: 0.63
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
Measured pKd: 5.05


Experiment series 10

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: PDZRN4
Protein B fragment: 214-318
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.7
Normalized holdup BI: 0.78
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: 5.38


Experiment series 11

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: PDZRN4
Protein B fragment: 214-318
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ

Measured dissociation constant (10-6M): 6
Standard deviation of dissociation constant (10-6M): 1.5
Experiment method: SPR
PUBMED ID of publication: 30726710
Number of measurements: 1
Measured pKd: 5.22