Details of experiments between P06463 and Q9UPQ7
Note that the results of all experiments are listed, regardless of the modification states of the fragments.



Experiment series 1

Protein A protein: HPV18-E6
Protein A fragment: 149-158
Protein A site: PBM
Protein A sequence: RLQRRRETQV
Protein A construct: biotin-ttds-RLQRRRETQV

Protein B protein: PDZRN3
Protein B fragment: 245-339
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.51
Holdup BI standard deviation: 0.15
Immobilized partner concentration in holdup experiment (10-6M): 13.3
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: 2
Measured pKd: 4.96


Experiment series 2

Protein A protein: HPV18-E6
Protein A fragment: 149-158
Protein A site: PBM
Protein A sequence: RLQRRRETQV
Protein A construct: biotin-ttds-RLQRRRETQV

Protein B protein: PDZRN3
Protein B fragment: 245-339
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ

Measured dissociation constant (10-6M): 13.75
Standard deviation of dissociation constant (10-6M): 0.83
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit

PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 4.86


Experiment series 3

Protein A protein: HPV18-E6
Protein A fragment: 149-158
Protein A site: PBM
Protein A modification: (TPO)156
Protein A sequence: RLQRRRETQV
Protein A construct: Biotin-ttds-RLQRRREpTQV

Protein B protein: PDZRN3
Protein B fragment: 245-339
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: -0.01
Normalized holdup BI: -0.01
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
The affinity was below the detection threshold of the assay.


Experiment series 4

Protein A protein: HPV18-E6
Protein A fragment: 149-158
Protein A site: PBM
Protein A sequence: RLQRRRETQV
Protein A construct: biotin-ttds-RLQRRRETQV

Protein B protein: PDZRN3
Protein B fragment: 245-339
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.68
Normalized holdup BI: 0.73
Immobilized partner concentration in holdup experiment (10-6M): 43
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: 4.82


Experiment series 5

Protein A protein: HPV18-E6
Protein A fragment: 149-158
Protein A site: PBM
Protein A sequence: RLQRRRETQV
Protein A construct: biotin-ttds-RLQRRRETQV

Protein B protein: PDZRN3
Protein B fragment: 245-339
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.49
Normalized holdup BI: 0.61
Immobilized partner concentration in holdup experiment (10-6M): 43
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: 4.58


Experiment series 6

Protein A protein: HPV18-E6
Protein A fragment: 149-158
Protein A site: PBM
Protein A sequence: RLQRRRETQV
Protein A construct: biotin-ttds-RLQRRRETQV

Protein B protein: PDZRN3
Protein B fragment: 245-339
Protein B site: PDZ_1
Protein B construct: his6-MBP-TEVsite-PDZ

Average holdup BI: 0.53
Normalized holdup BI: 0.63
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: 5.03