Details of experiments between the fragments Q07157~184-264 and P06463~149-158
Note that the results of all experiments are listed, regardless of the modification states of the fragments.



Experiment series 1

Protein A protein: TJP1
Protein A fragment: 184-264
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ

Protein B protein: HPV18-E6
Protein B fragment: 149-158
Protein B site: PBM
Protein B sequence: RLQRRRETQV
Protein B construct: biotin-ttds-RLQRRRETQV

Average holdup BI: 0.05
Holdup BI standard deviation: 0.02
Immobilized partner concentration in holdup experiment (10-6M): 13.3
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: 2
The affinity was below the detection threshold of the assay.


Experiment series 2

Protein A protein: TJP1
Protein A fragment: 184-264
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ

Protein B protein: HPV18-E6
Protein B fragment: 149-158
Protein B site: PBM
Protein B modification: (TPO)156
Protein B sequence: RLQRRRETQV
Protein B construct: Biotin-ttds-RLQRRREpTQV

Average holdup BI: 0
Normalized holdup BI: 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: 1
The affinity was below the detection threshold of the assay.


Experiment series 3

Protein A protein: TJP1
Protein A fragment: 184-264
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ

Protein B protein: HPV18-E6
Protein B fragment: 149-158
Protein B site: PBM
Protein B sequence: RLQRRRETQV
Protein B construct: biotin-ttds-RLQRRRETQV

Average holdup BI: -0.03
Normalized holdup BI: -0.04
Immobilized partner concentration in holdup experiment (10-6M): 43
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 4

Protein A protein: TJP1
Protein A fragment: 184-264
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ

Protein B protein: HPV18-E6
Protein B fragment: 149-158
Protein B site: PBM
Protein B sequence: RLQRRRETQV
Protein B construct: biotin-ttds-RLQRRRETQV

Average holdup BI: 0.03
Normalized holdup BI: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 43
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 5

Protein A protein: TJP1
Protein A fragment: 184-264
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ

Protein B protein: HPV18-E6
Protein B fragment: 149-158
Protein B site: PBM
Protein B sequence: RLQRRRETQV
Protein B construct: biotin-ttds-RLQRRRETQV

Average holdup BI: -0.06
Normalized holdup BI: -0.08
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
The affinity was below the detection threshold of the assay.