Details of experiments between the fragments Q15910~341-355 and O00499~513-593
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: EZH2
Protein A fragment: 341-355
Protein A site: PxxP_1
Protein A sequence: ERIKTPPKRPGGRRR
Protein A construct: Biotinylated 15-mer peptide (crude)
Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: 0.44
Holdup BI standard deviation: 0.01
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.06.29.
Number of measurements: 2
Measured pKd: 4.68
Experiment series 2
Protein A protein: EZH2
Protein A fragment: 341-355
Protein A site: PxxP_1
Protein A sequence: ERIKTPPKRPGGRRR
Protein A construct: Biotinylated 15-mer peptide (crude)
Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: T532M
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: 0.36
Holdup BI standard deviation: 0.01
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.06.29.
Number of measurements: 2
Measured pKd: 4.54
Experiment series 3
Protein A protein: EZH2
Protein A fragment: 341-355
Protein A site: PxxP_1
Protein A sequence: ERIKTPPKRPGGRRR
Protein A construct: Biotinylated 15-mer peptide (crude)
Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: D537V
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: 0.1
Holdup BI standard deviation: 0.07
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.10.24.
Number of measurements: 2
Measured pKd: 3.82
Experiment series 4
Protein A protein: EZH2
Protein A fragment: 341-355
Protein A site: PxxP_1
Protein A sequence: ERIKTPPKRPGGRRR
Protein A construct: Biotinylated 15-mer peptide (crude)
Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: F584S
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: 0.04
Holdup BI standard deviation: 0.02
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.10.24.
Number of measurements: 2
The affinity was below the detection threshold of the assay.
Experiment series 5
Protein A protein: EZH2
Protein A fragment: 341-355
Protein A site: PxxP_1
Protein A sequence: ERIKTPPKRPGGRRR
Protein A construct: Biotinylated 15-mer peptide (crude)
Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: P551L
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: 0.18
Holdup BI standard deviation: 0.04
Immobilized partner concentration in holdup experiment (10-6M): 6
Experiment method: HU (DAPF)
Details of affinity fitting: Bait concentration was adjusted to fit the profile to WT
Date of measurement: 2022.10.24.
Number of measurements: 2
Measured pKd: 4.59
Experiment series 6
Protein A protein: EZH2
Protein A fragment: 341-355
Protein A site: PxxP_1
Protein A sequence: ERIKTPPKRPGGRRR
Protein A construct: Biotinylated 15-mer peptide (crude)
Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: Q540H
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: 0.41
Holdup BI standard deviation: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.10.24.
Number of measurements: 2
Measured pKd: 4.62
Experiment series 7
Protein A protein: EZH2
Protein A fragment: 341-355
Protein A site: PxxP_1
Protein A sequence: ERIKTPPKRPGGRRR
Protein A construct: Biotinylated 15-mer peptide (crude)
Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: T532M
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: 0.32
Holdup BI standard deviation: 0.04
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.10.24.
Number of measurements: 2
Measured pKd: 4.45
Experiment series 8
Protein A protein: EZH2
Protein A fragment: 341-355
Protein A site: PxxP_1
Protein A sequence: ERIKTPPKRPGGRRR
Protein A construct: Biotinylated 15-mer peptide (crude)
Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: V566M
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: 0.33
Holdup BI standard deviation: 0.04
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.10.24.
Number of measurements: 2
Measured pKd: 4.48
Experiment series 9
Protein A protein: EZH2
Protein A fragment: 341-355
Protein A site: PxxP_1
Protein A sequence: ERIKTPPKRPGGRRR
Protein A construct: Biotinylated 15-mer peptide (crude)
Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: V583I
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: 0.2
Holdup BI standard deviation: 0.08
Immobilized partner concentration in holdup experiment (10-6M): 9
Experiment method: HU (DAPF)
Details of affinity fitting: Bait concentration was adjusted to fit the profile to WT
Date of measurement: 2022.10.24.
Number of measurements: 2
Measured pKd: 4.5
Experiment series 10
Protein A protein: EZH2
Protein A fragment: 341-355
Protein A site: PxxP_1
Protein A sequence: ERIKTPPKRPGGRRR
Protein A construct: Biotinylated 15-mer peptide (crude)
Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: 0.33
Holdup BI standard deviation: 0.04
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.10.24.
Number of measurements: 2
Measured pKd: 4.47
Experiment series 11
Protein A protein: EZH2
Protein A fragment: 341-355
Protein A site: PxxP_1
Protein A sequence: ERIKTPPKRPGGRRR
Protein A construct: Biotinylated 15-mer peptide (crude)
Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: Y531S
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: 0.17
Holdup BI standard deviation: 0.07
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.10.24.
Number of measurements: 2
Measured pKd: 4.07
Experiment series 12
Protein A protein: EZH2
Protein A fragment: 341-355
Protein A site: PxxP_1
Protein A sequence: ERIKTPPKRPGGRRR
Protein A construct: Biotinylated 15-mer peptide (crude)
Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: R581C
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: 0.19
Holdup BI standard deviation: 0.04
Immobilized partner concentration in holdup experiment (10-6M): 6
Experiment method: HU (DAPF)
Details of affinity fitting: Bait concentration was adjusted to fit the profile to WT
Date of measurement: 2022.10.24.
Number of measurements: 2
Measured pKd: 4.63
Experiment series 13
Protein A protein: EZH2
Protein A fragment: 341-355
Protein A site: PxxP_1
Protein A sequence: ERIKTPPKRPGGRRR
Protein A construct: Bio E E Ado Ado-ERIKTPPKRPGGRRR-CONH2
Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/16
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 14
Protein A protein: EZH2
Protein A fragment: 341-355
Protein A site: PxxP_1
Protein A sequence: ERIKTPPKRPGGRRR
Protein A construct: Bio E E Ado Ado-ERIKTPPKRPGGRRR-CONH2
Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: T532M
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/16
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 15
Protein A protein: EZH2
Protein A fragment: 341-355
Protein A site: PxxP_1
Protein A sequence: ERIKTPPKRPGGRRR
Protein A construct: Bio E E Ado Ado-ERIKTPPKRPGGRRR-CONH2
Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: Q540H
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.02
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/17
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 16
Protein A protein: EZH2
Protein A fragment: 341-355
Protein A site: PxxP_1
Protein A sequence: ERIKTPPKRPGGRRR
Protein A construct: Bio E E Ado Ado-ERIKTPPKRPGGRRR-CONH2
Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: D537V
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: -0
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/17
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 17
Protein A protein: EZH2
Protein A fragment: 341-355
Protein A site: PxxP_1
Protein A sequence: ERIKTPPKRPGGRRR
Protein A construct: Bio E E Ado Ado-ERIKTPPKRPGGRRR-CONH2
Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: V583I
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0
Immobilized partner concentration in holdup experiment (10-6M): 13
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/17
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 18
Protein A protein: EZH2
Protein A fragment: 341-355
Protein A site: PxxP_1
Protein A sequence: ERIKTPPKRPGGRRR
Protein A construct: Bio E E Ado Ado-ERIKTPPKRPGGRRR-CONH2
Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: P551L
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: -0.01
Immobilized partner concentration in holdup experiment (10-6M): 8
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/17
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 19
Protein A protein: EZH2
Protein A fragment: 341-355
Protein A site: PxxP_1
Protein A sequence: ERIKTPPKRPGGRRR
Protein A construct: Bio E E Ado Ado-ERIKTPPKRPGGRRR-CONH2
Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: F584S
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: -0.05
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/17
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 20
Protein A protein: EZH2
Protein A fragment: 341-355
Protein A site: PxxP_1
Protein A sequence: ERIKTPPKRPGGRRR
Protein A construct: Bio E E Ado Ado-ERIKTPPKRPGGRRR-CONH2
Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: Y531S
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/18
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 21
Protein A protein: EZH2
Protein A fragment: 341-355
Protein A site: PxxP_1
Protein A sequence: ERIKTPPKRPGGRRR
Protein A construct: Bio E E Ado Ado-ERIKTPPKRPGGRRR-CONH2
Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: R581C
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 10
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/18
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 22
Protein A protein: EZH2
Protein A fragment: 341-355
Protein A site: PxxP_1
Protein A sequence: ERIKTPPKRPGGRRR
Protein A construct: Bio E E Ado Ado-ERIKTPPKRPGGRRR-CONH2
Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: V566M
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/18
Number of measurements: 1
The affinity was below the detection threshold of the assay.