Details of experiments between the fragments P27228~140-149 and P78352~155-249
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: DLG4
Protein B fragment: 155-249
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.89
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: 5.36


Experiment series 2

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: DLG4
Protein B fragment: 155-249
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.05
Normalized holdup BI: 0.06
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: 3.56


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: DLG4
Protein B fragment: 155-249
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.64
Holdup BI standard deviation: 0.01
Normalized holdup BI: 0.83
Normalized holdup BI standard deviation: 0.01
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: 5.51


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: DLG4
Protein B fragment: 155-249
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.65
Normalized holdup BI: 0.78
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: 5.37


Experiment series 5

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: DLG4
Protein B fragment: 155-249
Protein B site: PDZ_2
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.68
Normalized holdup BI: 0.85
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: 5.58