Details of experiments between Q6P0Q8 and P60484
Note that the results of all experiments are listed, regardless of the modification states of the fragments.



Experiment series 1

Protein A protein: MAST2
Protein A fragment: 1102-1193
Protein A site: PDZ
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: 0.86
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: 5.37


Experiment series 2

Protein A protein: MAST2
Protein A fragment: 1102-1193
Protein A site: PDZ
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: 0.71
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: 4.87


Experiment series 3

Protein A protein: MAST2
Protein A fragment: 1102-1193
Protein A site: PDZ
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

Measured dissociation constant (10-6M): 3.14
Standard deviation of dissociation constant (10-6M): 0.18
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit

PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 5.5


Experiment series 4

Protein A protein: MAST2
Protein A fragment: 1102-1193
Protein A site: PDZ
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

Measured dissociation constant (10-6M): 2.56
Standard deviation of dissociation constant (10-6M): 0.13
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit

PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 5.59


Experiment series 5

Protein A protein: MAST2
Protein A fragment: 1102-1193
Protein A site: PDZ
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: 0.66
Normalized holdup BI: 0.66
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: 5.14


Experiment series 6

Protein A protein: MAST2
Protein A fragment: 1102-1193
Protein A site: PDZ
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: 0.74
Normalized holdup BI: 0.74
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: 5.24


Experiment series 7

Protein A protein: MAST2
Protein A fragment: 1102-1193
Protein A site: PDZ
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: 0.77
Normalized holdup BI: 0.85
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: 5.61


Experiment series 8

Protein A protein: MAST2
Protein A fragment: 1102-1193
Protein A site: PDZ
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: 0.82
Normalized holdup BI: 0.82
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: 5.53


Experiment series 9

Protein A protein: MAST2
Protein A fragment: 1102-1193
Protein A site: PDZ
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: 0.89
Normalized holdup BI: 0.89
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: 5.77


Experiment series 10

Protein A protein: MAST2
Protein A fragment: 1102-1193
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ

Protein B protein: PTEN
Protein B fragment: 394-403
Protein B site: PBM
Protein B sequence: EDQHTQITKV
Protein B construct: Biotin-ado-ado-EDQHTQITKV

Average holdup BI: 0.89
Normalized holdup BI: 0.89
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: Crude peptide synthesis product. The peptide concentration in the affinity conversion step may differ from reality. Not reported internal standards of holdup layout #6
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 5.75


Experiment series 11

Protein A protein: MAST2
Protein A fragment: 1102-1193
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ

Protein B protein: PTEN
Protein B fragment: 391-403
Protein B site: PBM
Protein B sequence: EDQHTQITKV
Protein B construct: Biotin-ttds-PFDEDQHTQITKV

Average holdup BI: 0.88
Immobilized partner concentration in holdup experiment (10-6M): 17
Experiment method: CALIP_holdup
Details of affinity fitting: lysozyme internal standard
PUBMED ID of publication: 33382810
Number of measurements: 1
Measured pKd: 5.74