Details of experiments between the fragments O00499~513-593 and Q6PJT7~329-343
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: BIN1
Protein A fragment: 513-593
Protein A site: SH3
Protein A construct: avi(biotin)-his6-MBP-TEVsite-SH3
Protein B protein: ZC3H14
Protein B fragment: 329-343
Protein B site: PxxP_1
Protein B sequence: SSSVSVPAKPERRPS
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.13
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: 3.92
Experiment series 2
Protein A protein: BIN1
Protein A fragment: 513-593
Protein A site: SH3
Protein A modification: T532M
Protein A construct: avi(biotin)-his6-MBP-TEVsite-SH3
Protein B protein: ZC3H14
Protein B fragment: 329-343
Protein B site: PxxP_1
Protein B sequence: SSSVSVPAKPERRPS
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.08
Holdup BI standard deviation: 0.03
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: 3.7
Experiment series 3
Protein A protein: BIN1
Protein A fragment: 513-593
Protein A site: SH3
Protein A modification: D537V
Protein A construct: avi(biotin)-his6-MBP-TEVsite-SH3
Protein B protein: ZC3H14
Protein B fragment: 329-343
Protein B site: PxxP_1
Protein B sequence: SSSVSVPAKPERRPS
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0
Holdup BI standard deviation: 0.01
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 4
Protein A protein: BIN1
Protein A fragment: 513-593
Protein A site: SH3
Protein A modification: F584S
Protein A construct: avi(biotin)-his6-MBP-TEVsite-SH3
Protein B protein: ZC3H14
Protein B fragment: 329-343
Protein B site: PxxP_1
Protein B sequence: SSSVSVPAKPERRPS
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: -0.04
Holdup BI standard deviation: 0.08
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: BIN1
Protein A fragment: 513-593
Protein A site: SH3
Protein A modification: P551L
Protein A construct: avi(biotin)-his6-MBP-TEVsite-SH3
Protein B protein: ZC3H14
Protein B fragment: 329-343
Protein B site: PxxP_1
Protein B sequence: SSSVSVPAKPERRPS
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.04
Holdup BI standard deviation: 0.01
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
The affinity was below the detection threshold of the assay.
Experiment series 6
Protein A protein: BIN1
Protein A fragment: 513-593
Protein A site: SH3
Protein A modification: Q540H
Protein A construct: avi(biotin)-his6-MBP-TEVsite-SH3
Protein B protein: ZC3H14
Protein B fragment: 329-343
Protein B site: PxxP_1
Protein B sequence: SSSVSVPAKPERRPS
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.08
Holdup BI standard deviation: 0.06
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: 3.7
Experiment series 7
Protein A protein: BIN1
Protein A fragment: 513-593
Protein A site: SH3
Protein A modification: T532M
Protein A construct: avi(biotin)-his6-MBP-TEVsite-SH3
Protein B protein: ZC3H14
Protein B fragment: 329-343
Protein B site: PxxP_1
Protein B sequence: SSSVSVPAKPERRPS
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.07
Holdup BI standard deviation: 0
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: 3.63
Experiment series 8
Protein A protein: BIN1
Protein A fragment: 513-593
Protein A site: SH3
Protein A modification: V566M
Protein A construct: avi(biotin)-his6-MBP-TEVsite-SH3
Protein B protein: ZC3H14
Protein B fragment: 329-343
Protein B site: PxxP_1
Protein B sequence: SSSVSVPAKPERRPS
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.06
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
Measured pKd: 3.58
Experiment series 9
Protein A protein: BIN1
Protein A fragment: 513-593
Protein A site: SH3
Protein A modification: V583I
Protein A construct: avi(biotin)-his6-MBP-TEVsite-SH3
Protein B protein: ZC3H14
Protein B fragment: 329-343
Protein B site: PxxP_1
Protein B sequence: SSSVSVPAKPERRPS
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.01
Holdup BI standard deviation: 0.02
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
The affinity was below the detection threshold of the assay.
Experiment series 10
Protein A protein: BIN1
Protein A fragment: 513-593
Protein A site: SH3
Protein A construct: avi(biotin)-his6-MBP-TEVsite-SH3
Protein B protein: ZC3H14
Protein B fragment: 329-343
Protein B site: PxxP_1
Protein B sequence: SSSVSVPAKPERRPS
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.09
Holdup BI standard deviation: 0.01
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: 3.76
Experiment series 11
Protein A protein: BIN1
Protein A fragment: 513-593
Protein A site: SH3
Protein A modification: Y531S
Protein A construct: avi(biotin)-his6-MBP-TEVsite-SH3
Protein B protein: ZC3H14
Protein B fragment: 329-343
Protein B site: PxxP_1
Protein B sequence: SSSVSVPAKPERRPS
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.21
Holdup BI standard deviation: 0
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: 4.2
Experiment series 12
Protein A protein: BIN1
Protein A fragment: 513-593
Protein A site: SH3
Protein A modification: R581C
Protein A construct: avi(biotin)-his6-MBP-TEVsite-SH3
Protein B protein: ZC3H14
Protein B fragment: 329-343
Protein B site: PxxP_1
Protein B sequence: SSSVSVPAKPERRPS
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.03
Holdup BI standard deviation: 0.01
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
The affinity was below the detection threshold of the assay.
Experiment series 13
Protein A protein: BIN1
Protein A fragment: 513-593
Protein A site: SH3
Protein A construct: his6-MBP-TEVsite-SH3
Protein B protein: ZC3H14
Protein B fragment: 329-343
Protein B site: PxxP_1
Protein B sequence: SSSVSVPAKPERRPS
Protein B construct: Bio Ado Ado-SSSVSVPAKPERRPS-CONH2
Average holdup BI: 0.08
Holdup BI standard deviation: 0.01
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: 2
Measured pKd: 3.72
Experiment series 14
Protein A protein: BIN1
Protein A fragment: 513-593
Protein A site: SH3
Protein A modification: T532M
Protein A construct: his6-MBP-TEVsite-SH3
Protein B protein: ZC3H14
Protein B fragment: 329-343
Protein B site: PxxP_1
Protein B sequence: SSSVSVPAKPERRPS
Protein B construct: Bio Ado Ado-SSSVSVPAKPERRPS-CONH2
Average holdup BI: 0.09
Holdup BI standard deviation: 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: 2
Measured pKd: 3.75
Experiment series 15
Protein A protein: BIN1
Protein A fragment: 513-593
Protein A site: SH3
Protein A modification: Q540H
Protein A construct: his6-MBP-TEVsite-SH3
Protein B protein: ZC3H14
Protein B fragment: 329-343
Protein B site: PxxP_1
Protein B sequence: SSSVSVPAKPERRPS
Protein B construct: Bio Ado Ado-SSSVSVPAKPERRPS-CONH2
Average holdup BI: 0.09
Holdup BI standard deviation: 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: 2
Measured pKd: 3.74
Experiment series 16
Protein A protein: BIN1
Protein A fragment: 513-593
Protein A site: SH3
Protein A modification: D537V
Protein A construct: his6-MBP-TEVsite-SH3
Protein B protein: ZC3H14
Protein B fragment: 329-343
Protein B site: PxxP_1
Protein B sequence: SSSVSVPAKPERRPS
Protein B construct: Bio Ado Ado-SSSVSVPAKPERRPS-CONH2
Average holdup BI: -0.02
Holdup BI standard deviation: 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: 2023/01/17
Number of measurements: 2
The affinity was below the detection threshold of the assay.
Experiment series 17
Protein A protein: BIN1
Protein A fragment: 513-593
Protein A site: SH3
Protein A modification: V583I
Protein A construct: his6-MBP-TEVsite-SH3
Protein B protein: ZC3H14
Protein B fragment: 329-343
Protein B site: PxxP_1
Protein B sequence: SSSVSVPAKPERRPS
Protein B construct: Bio Ado Ado-SSSVSVPAKPERRPS-CONH2
Average holdup BI: 0.03
Holdup BI standard deviation: 0.03
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: 2
The affinity was below the detection threshold of the assay.
Experiment series 18
Protein A protein: BIN1
Protein A fragment: 513-593
Protein A site: SH3
Protein A modification: P551L
Protein A construct: his6-MBP-TEVsite-SH3
Protein B protein: ZC3H14
Protein B fragment: 329-343
Protein B site: PxxP_1
Protein B sequence: SSSVSVPAKPERRPS
Protein B construct: Bio Ado Ado-SSSVSVPAKPERRPS-CONH2
Average holdup BI: 0.04
Holdup BI standard deviation: 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: 2
The affinity was below the detection threshold of the assay.
Experiment series 19
Protein A protein: BIN1
Protein A fragment: 513-593
Protein A site: SH3
Protein A modification: F584S
Protein A construct: his6-MBP-TEVsite-SH3
Protein B protein: ZC3H14
Protein B fragment: 329-343
Protein B site: PxxP_1
Protein B sequence: SSSVSVPAKPERRPS
Protein B construct: Bio Ado Ado-SSSVSVPAKPERRPS-CONH2
Average holdup BI: -0.01
Holdup BI standard deviation: 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: 2023/01/17
Number of measurements: 2
The affinity was below the detection threshold of the assay.
Experiment series 20
Protein A protein: BIN1
Protein A fragment: 513-593
Protein A site: SH3
Protein A modification: Y531S
Protein A construct: his6-MBP-TEVsite-SH3
Protein B protein: ZC3H14
Protein B fragment: 329-343
Protein B site: PxxP_1
Protein B sequence: SSSVSVPAKPERRPS
Protein B construct: Bio Ado Ado-SSSVSVPAKPERRPS-CONH2
Average holdup BI: 0.23
Holdup BI standard deviation: 0.01
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: 2
Measured pKd: 4.25
Experiment series 21
Protein A protein: BIN1
Protein A fragment: 513-593
Protein A site: SH3
Protein A modification: R581C
Protein A construct: his6-MBP-TEVsite-SH3
Protein B protein: ZC3H14
Protein B fragment: 329-343
Protein B site: PxxP_1
Protein B sequence: SSSVSVPAKPERRPS
Protein B construct: Bio Ado Ado-SSSVSVPAKPERRPS-CONH2
Average holdup BI: 0
Holdup BI standard deviation: 0.02
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: 2
The affinity was below the detection threshold of the assay.
Experiment series 22
Protein A protein: BIN1
Protein A fragment: 513-593
Protein A site: SH3
Protein A modification: V566M
Protein A construct: his6-MBP-TEVsite-SH3
Protein B protein: ZC3H14
Protein B fragment: 329-343
Protein B site: PxxP_1
Protein B sequence: SSSVSVPAKPERRPS
Protein B construct: Bio Ado Ado-SSSVSVPAKPERRPS-CONH2
Average holdup BI: 0.1
Holdup BI standard deviation: 0.01
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: 2
Measured pKd: 3.78