Details of experiments between Q13424 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: SNTA1
Protein A fragment: 84-171
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: 0.41
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: 4.4


Experiment series 2

Protein A protein: SNTA1
Protein A fragment: 84-171
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: 0.18
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: 3.79


Experiment series 3

Protein A protein: SNTA1
Protein A fragment: 84-171
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): 193.98
Standard deviation of dissociation constant (10-6M): 17.47
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit

PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 3.71


Experiment series 4

Protein A protein: SNTA1
Protein A fragment: 84-171
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): 45.27
Standard deviation of dissociation constant (10-6M): 3.44
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit

PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 4.34


Experiment series 5

Protein A protein: SNTA1
Protein A fragment: 84-171
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: 0.15
Normalized holdup BI: 0.17
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: 4.04


Experiment series 6

Protein A protein: SNTA1
Protein A fragment: 84-171
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: 0.38
Normalized holdup BI: 0.44
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: 4.71


Experiment series 7

Protein A protein: SNTA1
Protein A fragment: 84-171
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: 0.32
Normalized holdup BI: 0.42
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: 4.68


Experiment series 8

Protein A protein: SNTA1
Protein A fragment: 84-171
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: 0.34
Normalized holdup BI: 0.38
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: 4.57


Experiment series 9

Protein A protein: SNTA1
Protein A fragment: 84-171
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: 0.14
Normalized holdup BI: 0.15
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: 4.02


Experiment series 10

Protein A protein: SNTA1
Protein A fragment: 84-171
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: 0.14
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: 4