Details of experiments between the fragments P60484~393-403 and Q9P202~813-904
Note that the results of all experiments are listed, regardless of the modification states of the fragments.



Experiment series 1

Protein A protein: PTEN
Protein A fragment: 393-403
Protein A site: PBM
Protein A modification: (ALY)402
Protein A sequence: EDQHTQITKV
Protein A construct: biotin-ttds-DEDQHTQITKacV

Protein B protein: DFNB31
Protein B fragment: 813-904
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.28
Immobilized partner concentration in holdup experiment (10-6M): 29
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: 4.13


Experiment series 2

Protein A protein: PTEN
Protein A fragment: 393-403
Protein A site: PBM
Protein A sequence: EDQHTQITKV
Protein A construct: biotin-ttds-DEDQHTQITKV

Protein B protein: DFNB31
Protein B fragment: 813-904
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.09
Immobilized partner concentration in holdup experiment (10-6M): 36
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: PTEN
Protein A fragment: 393-403
Protein A site: PBM
Protein A modification: (ALY)402
Protein A sequence: EDQHTQITKV
Protein A construct: biotin-ttds-DEDQHTQITKacV

Protein B protein: DFNB31
Protein B fragment: 813-904
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.22
Normalized holdup BI: 0.22
Immobilized partner concentration in holdup experiment (10-6M): 17
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: 4.25


Experiment series 4

Protein A protein: PTEN
Protein A fragment: 393-403
Protein A site: PBM
Protein A sequence: EDQHTQITKV
Protein A construct: biotin-ttds-DEDQHTQITKV

Protein B protein: DFNB31
Protein B fragment: 813-904
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.09
Normalized holdup BI: 0.09
Immobilized partner concentration in holdup experiment (10-6M): 19
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: 3.71


Experiment series 5

Protein A protein: PTEN
Protein A fragment: 393-403
Protein A site: PBM
Protein A modification: (ALY)402
Protein A sequence: EDQHTQITKV
Protein A construct: biotin-ttds-DEDQHTQITKacV

Protein B protein: DFNB31
Protein B fragment: 813-904
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.22
Normalized holdup BI: 0.22
Immobilized partner concentration in holdup experiment (10-6M): 17
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: 4.23


Experiment series 6

Protein A protein: PTEN
Protein A fragment: 393-403
Protein A site: PBM
Protein A modification: (ALY)402
Protein A sequence: EDQHTQITKV
Protein A construct: biotin-ttds-DEDQHTQITKacV

Protein B protein: DFNB31
Protein B fragment: 813-904
Protein B site: PDZ_3
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.31
Normalized holdup BI: 0.31
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: 4.42