Details of experiments between the fragments Q6P0Q8~1102-1193 and Q81964~151-160
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: HPV59-E6
Protein B fragment: 151-160
Protein B site: PBM
Protein B sequence: QRQARSETLV
Protein B construct: biotin-ttds-QRQARSETLV

Measured dissociation constant (10-6M): 12.7
Standard deviation of dissociation constant (10-6M): 0.74
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit

PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 4.9


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: HPV59-E6
Protein B fragment: 151-160
Protein B site: PBM
Protein B sequence: QRQARSETLV
Protein B construct: biotin-ttds-QRQARSETLV

Average holdup BI: 0.29
Immobilized partner concentration in holdup experiment (10-6M): 7
Experiment method: SAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normal layout
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 4.85


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: HPV59-E6
Protein B fragment: 151-160
Protein B site: PBM
Protein B sequence: QRQARSETLV
Protein B construct: biotin-ttds-QRQARSETLV

Average holdup BI: 0.55
Holdup BI standard deviation: 0.01
Normalized holdup BI: 0.55
Normalized holdup BI standard deviation: 0.01
Immobilized partner concentration in holdup experiment (10-6M): 13.7
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: 5.03