Details of experiments between Q7Z6J2 and Q5SYB0
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: GRASP
Protein A fragment: 386-395
Protein A site: PBM
Protein A sequence: RSLEEEESQL
Protein A construct: biotin-ttds-RSLEEEESQL
Protein B protein: FRMPD1
Protein B fragment: 53-142
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.02
Immobilized partner concentration in holdup experiment (10-6M): 10
Experiment method: SAPF_holdup
Details of affinity fitting: fluorescein + mCherry internal standards + normal layout
PUBMED ID of publication: 36115835
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 2
Protein A protein: GRASP
Protein A fragment: 386-395
Protein A site: PBM
Protein A sequence: RSLEEEESQL
Protein A construct: biotin-ttds-RSLEEEESQL
Protein B protein: FRMPD1
Protein B fragment: 53-142
Protein B site: PDZ
Protein B construct: his6-MBP-TEVsite-PDZ
Average holdup BI: 0.08
Normalized holdup BI: 0.08
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
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 3.71
Experiment series 3
Protein A protein: GRASP
Protein A fragment: 96-193
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: FRMPD1
Protein B fragment: 1569-1578
Protein B site: PBM
Protein B sequence: TQKFRASTAL
Protein B construct: Biotin-Ahx-TQKFRASTAL
Average holdup BI: 0.38
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
PUBMED ID of publication: 36115835
Number of measurements: 1
Measured pKd: 4.56
Experiment series 4
Protein A protein: GRASP
Protein A fragment: 96-193
Protein A site: PDZ
Protein A construct: his6-MBP-TEVsite-PDZ
Protein B protein: FRMPD1
Protein B fragment: 1569-1578
Protein B site: PBM
Protein B sequence: TQKFRASTAL
Protein B construct: Biotin-Ahx-TQKFRASTAL
Average holdup BI: 0.33
Normalized holdup BI: 0.33
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: 4.46