Details of experiments between P27228 and Q9H5P4
Note that the results of all experiments are listed, regardless of the modification states of the fragments.



Experiment series 1

Protein A protein: HPV35-E6
Protein A fragment: 140-149
Protein A site: PBM
Protein A sequence: SKPTRRETEV
Protein A construct: biotin-ttds-SKPTRRETEV

Protein B protein: PDZD7
Protein B fragment: 858-953
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.31
Immobilized partner concentration in holdup experiment (10-6M): 39.7
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: 4.07


Experiment series 2

Protein A protein: HPV35-E6
Protein A fragment: 140-149
Protein A site: PBM
Protein A sequence: SKPTRRETEV
Protein A construct: biotin-ttds-SKPTRRETEV

Protein B protein: PDZD7
Protein B fragment: 82-170
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): 39.7
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: HPV35-E6
Protein A fragment: 140-149
Protein A site: PBM
Protein A sequence: SKPTRRETEV
Protein A construct: biotin-ttds-SKPTRRETEV

Protein B protein: PDZD7
Protein B fragment: 199-313
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: -0.1
Immobilized partner concentration in holdup experiment (10-6M): 39.7
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 4

Protein A protein: HPV35-E6
Protein A fragment: 140-149
Protein A site: PBM
Protein A sequence: SKPTRRETEV
Protein A construct: biotin-ttds-SKPTRRETEV

Protein B protein: PDZD7
Protein B fragment: 858-953
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ

Measured dissociation constant (10-6M): 35.01
Standard deviation of dissociation constant (10-6M): 1.93
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit

PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 4.46


Experiment series 5

Protein A protein: HPV35-E6
Protein A fragment: 140-149
Protein A site: PBM
Protein A modification: (TPO)147
Protein A sequence: SKPTRRETEV
Protein A construct: Biotin-ttds-SKPTRREpTEV

Protein B protein: PDZD7
Protein B fragment: 858-953
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.06
Normalized holdup BI: 0.07
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: 3.61


Experiment series 6

Protein A protein: HPV35-E6
Protein A fragment: 140-149
Protein A site: PBM
Protein A sequence: SKPTRRETEV
Protein A construct: biotin-ttds-SKPTRRETEV

Protein B protein: PDZD7
Protein B fragment: 858-953
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.21
Holdup BI standard deviation: 0.02
Normalized holdup BI: 0.26
Normalized holdup BI standard deviation: 0.02
Immobilized partner concentration in holdup experiment (10-6M): 18.55
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: 4.3


Experiment series 7

Protein A protein: HPV35-E6
Protein A fragment: 140-149
Protein A site: PBM
Protein A sequence: SKPTRRETEV
Protein A construct: biotin-ttds-SKPTRRETEV

Protein B protein: PDZD7
Protein B fragment: 858-953
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.21
Normalized holdup BI: 0.26
Immobilized partner concentration in holdup experiment (10-6M): 18.55
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: 4.3


Experiment series 8

Protein A protein: HPV35-E6
Protein A fragment: 140-149
Protein A site: PBM
Protein A sequence: SKPTRRETEV
Protein A construct: biotin-ttds-SKPTRRETEV

Protein B protein: PDZD7
Protein B fragment: 858-953
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.24
Normalized holdup BI: 0.26
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 #6
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 4.31