Details of experiments between Q14160 and P27037
Note that the results of all experiments are listed, regardless of the modification states of the fragments.



Experiment series 1

Protein A protein: SCRIB
Protein A fragment: 718-814
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ

Protein B protein: ACVR2A
Protein B fragment: 504-513
Protein B site: PBM
Protein B sequence: VDFPPKESSL
Protein B construct: biotin-ttds-VDFPPKESSL

Measured dissociation constant (10-6M): 85.41
Standard deviation of dissociation constant (10-6M): 2.82
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit

PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 4.07


Experiment series 2

Protein A protein: SCRIB
Protein A fragment: 999-1093
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ

Protein B protein: ACVR2A
Protein B fragment: 504-513
Protein B site: PBM
Protein B sequence: VDFPPKESSL
Protein B construct: biotin-ttds-VDFPPKESSL

Average holdup BI: 0.63
Normalized holdup BI: 0.7
Immobilized partner concentration in holdup experiment (10-6M): 33
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: 4.88


Experiment series 3

Protein A protein: SCRIB
Protein A fragment: 718-814
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ

Protein B protein: ACVR2A
Protein B fragment: 504-513
Protein B site: PBM
Protein B sequence: VDFPPKESSL
Protein B construct: biotin-ttds-VDFPPKESSL

Average holdup BI: 0.17
Holdup BI standard deviation: 0.08
Normalized holdup BI: 0.18
Normalized holdup BI standard deviation: 0.08
Immobilized partner concentration in holdup experiment (10-6M): 33
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: 4
Measured pKd: 3.83


Experiment series 4

Protein A protein: SCRIB
Protein A fragment: 858-949
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ

Protein B protein: ACVR2A
Protein B fragment: 504-513
Protein B site: PBM
Protein B sequence: VDFPPKESSL
Protein B construct: biotin-ttds-VDFPPKESSL

Average holdup BI: 0.07
Normalized holdup BI: 0.09
Immobilized partner concentration in holdup experiment (10-6M): 33
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: 3.48