Details of experiments between Q86UL8 and Q81964
Note that the results of all experiments are listed, regardless of the modification states of the fragments.



Experiment series 1

Protein A protein: MAGI2
Protein A fragment: 410-522
Protein A site: PDZ_2
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): 0.5
Standard deviation of dissociation constant (10-6M): 0.04
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit

PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 6.3


Experiment series 2

Protein A protein: MAGI2
Protein A fragment: 772-865
Protein A site: PDZ_4
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: 0.82
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: 6.1


Experiment series 3

Protein A protein: MAGI2
Protein A fragment: 410-522
Protein A site: PDZ_2
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: 0.77
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: 5.92


Experiment series 4

Protein A protein: MAGI2
Protein A fragment: 915-1017
Protein A site: PDZ_5
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: 0.42
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: 5.14


Experiment series 5

Protein A protein: MAGI2
Protein A fragment: 600-682
Protein A site: PDZ_3
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: 0.12
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: 4.33


Experiment series 6

Protein A protein: MAGI2
Protein A fragment: 1-102
Protein A site: PDZ_1
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: -0.1
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
The affinity was below the detection threshold of the assay.


Experiment series 7

Protein A protein: MAGI2
Protein A fragment: 1141-1229
Protein A site: PDZ_6
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: -0.01
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
The affinity was below the detection threshold of the assay.


Experiment series 8

Protein A protein: MAGI2
Protein A fragment: 410-522
Protein A site: PDZ_2
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: 0.84
Holdup BI standard deviation: 0.09
Normalized holdup BI: 0.92
Normalized holdup BI standard deviation: 0.1
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: 6.07


Experiment series 9

Protein A protein: MAGI2
Protein A fragment: 772-865
Protein A site: PDZ_4
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: 0.6
Normalized holdup BI: 0.84
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: 1
Measured pKd: 5.71


Experiment series 10

Protein A protein: MAGI2
Protein A fragment: 915-1017
Protein A site: PDZ_5
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
Normalized holdup BI: 0.55
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: 1
Measured pKd: 5.02


Experiment series 11

Protein A protein: MAGI2
Protein A fragment: 600-682
Protein A site: PDZ_3
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: 0.33
Normalized holdup BI: 0.39
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: 1
Measured pKd: 4.73


Experiment series 12

Protein A protein: MAGI2
Protein A fragment: 1141-1229
Protein A site: PDZ_6
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: 0.06
Normalized holdup BI: 0.07
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: 1
Measured pKd: 3.74