Details of experiments between the fragments P60484~393-403 and Q13884~107-196
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: SNTB1
Protein B fragment: 107-196
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.55
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: 4.66


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: SNTB1
Protein B fragment: 107-196
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.16
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: 3.72


Experiment series 3

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: SNTB1
Protein B fragment: 107-196
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ

Measured dissociation constant (10-6M): 67.04
Standard deviation of dissociation constant (10-6M): 45.34
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit

PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 4.17


Experiment series 4

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: SNTB1
Protein B fragment: 107-196
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ

Measured dissociation constant (10-6M): 14.53
Standard deviation of dissociation constant (10-6M): 1.79
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit

PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 4.84


Experiment series 5

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: SNTB1
Protein B fragment: 107-196
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.15
Holdup BI standard deviation: 0.09
Normalized holdup BI: 0.18
Normalized holdup BI standard deviation: 0.1
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: 2
Measured pKd: 4.08


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: SNTB1
Protein B fragment: 107-196
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.4
Normalized holdup BI: 0.48
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: 4.79


Experiment series 7

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: SNTB1
Protein B fragment: 107-196
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.36
Normalized holdup BI: 0.4
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: 4.63


Experiment series 8

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: SNTB1
Protein B fragment: 107-196
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.44
Normalized holdup BI: 0.49
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: 4.78