Details of experiments between the fragments Q12959~221-314 and P60484~393-403
Note that the results of all experiments are listed, regardless of the modification states of the fragments.



Experiment series 1

Protein A protein: DLG1
Protein A fragment: 221-314
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ

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

Average holdup BI: 0.17
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: 3.85


Experiment series 2

Protein A protein: DLG1
Protein A fragment: 221-314
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ

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

Average holdup BI: 0.11
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
Measured pKd: 3.54


Experiment series 3

Protein A protein: DLG1
Protein A fragment: 221-314
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ

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

Average holdup BI: 0.16
Normalized holdup BI: 0.19
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: 4.15


Experiment series 4

Protein A protein: DLG1
Protein A fragment: 221-314
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ

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

Average holdup BI: 0.06
Normalized holdup BI: 0.08
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: 3.64


Experiment series 5

Protein A protein: DLG1
Protein A fragment: 221-314
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ

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

Average holdup BI: 0.18
Normalized holdup BI: 0.23
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: 4.28


Experiment series 6

Protein A protein: DLG1
Protein A fragment: 221-314
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ

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

Average holdup BI: 0.13
Normalized holdup BI: 0.16
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: 4.03