Details of experiments between P78352 and P27228
Note that the results of all experiments are listed, regardless of the modification states of the fragments.



Experiment series 1

Protein A protein: DLG4
Protein A fragment: 155-249
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ

Protein B protein: HPV35-E6
Protein B fragment: 140-149
Protein B site: PBM
Protein B sequence: SKPTRRETEV
Protein B construct: biotin-ttds-SKPTRRETEV

Average holdup BI: 0.89
Immobilized partner concentration in holdup experiment (10-6M): 39.7
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: 1
Measured pKd: 5.36


Experiment series 2

Protein A protein: DLG4
Protein A fragment: 306-403
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ

Protein B protein: HPV35-E6
Protein B fragment: 140-149
Protein B site: PBM
Protein B sequence: SKPTRRETEV
Protein B construct: biotin-ttds-SKPTRRETEV

Average holdup BI: 0.78
Immobilized partner concentration in holdup experiment (10-6M): 39.7
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: 1
Measured pKd: 4.98


Experiment series 3

Protein A protein: DLG4
Protein A fragment: 62-152
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ

Protein B protein: HPV35-E6
Protein B fragment: 140-149
Protein B site: PBM
Protein B sequence: SKPTRRETEV
Protein B construct: biotin-ttds-SKPTRRETEV

Average holdup BI: 0.76
Immobilized partner concentration in holdup experiment (10-6M): 39.7
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: 1
Measured pKd: 4.94


Experiment series 4

Protein A protein: DLG4
Protein A fragment: 306-403
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ

Protein B protein: HPV35-E6
Protein B fragment: 140-149
Protein B site: PBM
Protein B sequence: SKPTRRETEV
Protein B construct: biotin-ttds-SKPTRRETEV

Measured dissociation constant (10-6M): 6.37
Standard deviation of dissociation constant (10-6M): 0.25
Experiment method: competitive FP
Details of affinity fitting: Competitive binding equation in ProFit

PUBMED ID of publication: 36115835
Number of measurements: 3
Measured pKd: 5.2


Experiment series 5

Protein A protein: DLG4
Protein A fragment: 155-249
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ

Protein B protein: HPV35-E6
Protein B fragment: 140-149
Protein B site: PBM
Protein B modification: (TPO)147
Protein B sequence: SKPTRRETEV
Protein B construct: Biotin-ttds-SKPTRREpTEV

Average holdup BI: 0.05
Normalized holdup BI: 0.06
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: 3.56


Experiment series 6

Protein A protein: DLG4
Protein A fragment: 62-152
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ

Protein B protein: HPV35-E6
Protein B fragment: 140-149
Protein B site: PBM
Protein B modification: (TPO)147
Protein B sequence: SKPTRRETEV
Protein B construct: Biotin-ttds-SKPTRREpTEV

Average holdup BI: -0.09
Normalized holdup BI: -0.11
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 7

Protein A protein: DLG4
Protein A fragment: 306-403
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ

Protein B protein: HPV35-E6
Protein B fragment: 140-149
Protein B site: PBM
Protein B modification: (TPO)147
Protein B sequence: SKPTRRETEV
Protein B construct: Biotin-ttds-SKPTRREpTEV

Average holdup BI: 0.04
Normalized holdup BI: 0.04
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 8

Protein A protein: DLG4
Protein A fragment: 155-249
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ

Protein B protein: HPV35-E6
Protein B fragment: 140-149
Protein B site: PBM
Protein B sequence: SKPTRRETEV
Protein B construct: biotin-ttds-SKPTRRETEV

Average holdup BI: 0.64
Holdup BI standard deviation: 0.01
Normalized holdup BI: 0.83
Normalized holdup BI standard deviation: 0.01
Immobilized partner concentration in holdup experiment (10-6M): 18.55
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: 5.51


Experiment series 9

Protein A protein: DLG4
Protein A fragment: 306-403
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ

Protein B protein: HPV35-E6
Protein B fragment: 140-149
Protein B site: PBM
Protein B sequence: SKPTRRETEV
Protein B construct: biotin-ttds-SKPTRRETEV

Average holdup BI: 0.62
Holdup BI standard deviation: 0.01
Normalized holdup BI: 0.71
Normalized holdup BI standard deviation: 0.01
Immobilized partner concentration in holdup experiment (10-6M): 18.55
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: 3
Measured pKd: 5.2


Experiment series 10

Protein A protein: DLG4
Protein A fragment: 62-152
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ

Protein B protein: HPV35-E6
Protein B fragment: 140-149
Protein B site: PBM
Protein B sequence: SKPTRRETEV
Protein B construct: biotin-ttds-SKPTRRETEV

Average holdup BI: 0.55
Holdup BI standard deviation: 0
Normalized holdup BI: 0.66
Normalized holdup BI standard deviation: 0
Immobilized partner concentration in holdup experiment (10-6M): 18.55
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: 5.09


Experiment series 11

Protein A protein: DLG4
Protein A fragment: 62-152
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ

Protein B protein: HPV35-E6
Protein B fragment: 140-149
Protein B site: PBM
Protein B sequence: SKPTRRETEV
Protein B construct: biotin-ttds-SKPTRRETEV

Average holdup BI: 0.54
Normalized holdup BI: 0.54
Immobilized partner concentration in holdup experiment (10-6M): 18.55
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: 4.85


Experiment series 12

Protein A protein: DLG4
Protein A fragment: 155-249
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ

Protein B protein: HPV35-E6
Protein B fragment: 140-149
Protein B site: PBM
Protein B sequence: SKPTRRETEV
Protein B construct: biotin-ttds-SKPTRRETEV

Average holdup BI: 0.65
Normalized holdup BI: 0.78
Immobilized partner concentration in holdup experiment (10-6M): 18.55
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: 5.37


Experiment series 13

Protein A protein: DLG4
Protein A fragment: 306-403
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ

Protein B protein: HPV35-E6
Protein B fragment: 140-149
Protein B site: PBM
Protein B sequence: SKPTRRETEV
Protein B construct: biotin-ttds-SKPTRRETEV

Average holdup BI: 0.47
Normalized holdup BI: 0.59
Immobilized partner concentration in holdup experiment (10-6M): 18.55
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: 4.94


Experiment series 14

Protein A protein: DLG4
Protein A fragment: 306-403
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ

Protein B protein: HPV35-E6
Protein B fragment: 140-149
Protein B site: PBM
Protein B sequence: SKPTRRETEV
Protein B construct: biotin-ttds-SKPTRRETEV

Average holdup BI: 0.52
Normalized holdup BI: 0.59
Immobilized partner concentration in holdup experiment (10-6M): 18.55
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: 4.96


Experiment series 15

Protein A protein: DLG4
Protein A fragment: 306-403
Protein A site: PDZ_3
Protein A construct: his6-MBP-TEVsite-PDZ

Protein B protein: HPV35-E6
Protein B fragment: 140-149
Protein B site: PBM
Protein B sequence: SKPTRRETEV
Protein B construct: biotin-ttds-SKPTRRETEV

Average holdup BI: 0.7
Normalized holdup BI: 0.83
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: 5.51


Experiment series 16

Protein A protein: DLG4
Protein A fragment: 155-249
Protein A site: PDZ_2
Protein A construct: his6-MBP-TEVsite-PDZ

Protein B protein: HPV35-E6
Protein B fragment: 140-149
Protein B site: PBM
Protein B sequence: SKPTRRETEV
Protein B construct: biotin-ttds-SKPTRRETEV

Average holdup BI: 0.68
Normalized holdup BI: 0.85
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: 5.58


Experiment series 17

Protein A protein: DLG4
Protein A fragment: 62-152
Protein A site: PDZ_1
Protein A construct: his6-MBP-TEVsite-PDZ

Protein B protein: HPV35-E6
Protein B fragment: 140-149
Protein B site: PBM
Protein B sequence: SKPTRRETEV
Protein B construct: biotin-ttds-SKPTRRETEV

Average holdup BI: 0.54
Normalized holdup BI: 0.64
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: 5.06