Details of experiments between O00499 and Q96B97
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: SH3KBP1
Protein B fragment: 1-665
Protein B construct: endogenous protein (Jurkat)
Average holdup BI: 0.28
Immobilized partner concentration in holdup experiment (10-6M): 10
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 6.77
Experiment method: nHU_Jurkat
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.06.29.
Experimental details: -log10(P Two-sided unpaired T-test (6 vs 3))
Number of measurements: 3
Measured pKd: 4.58
Experiment series 2
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: SH3KBP1
Protein B fragment: 1-665
Protein B construct: endogenous protein (Jurkat)
Average holdup BI: 0.63
Immobilized partner concentration in holdup experiment (10-6M): 10
P value (usually -log10(P Two-sided unpaired T-test), but double check the experimental details): 3.1
Experiment method: nHU_Jurkat
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022.06.29.
Experimental details: -log10(P Two-sided unpaired T-test (6 vs 3))
Number of measurements: 3
Measured pKd: 5.23
Experiment series 3
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: SH3KBP1
Protein B fragment: 360-374
Protein B site: PxxP_1
Protein B sequence: EIKKIPPERPEMLPN
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: -0.02
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
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 construct: avi(biotin)-his6-MBP-TEVsite-SH3
Protein B protein: SH3KBP1
Protein B fragment: 390-404
Protein B site: PxxP_2
Protein B sequence: LQKPSVPAIPPKKPR
Protein B construct: Biotinylated 15-mer peptide (crude)
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 5
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: SH3KBP1
Protein B fragment: 413-427
Protein B site: PxxP_3
Protein B sequence: RPGALPPRRPERPVG
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.1
Holdup BI standard deviation: 0.04
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: 3.81
Experiment series 6
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: SH3KBP1
Protein B fragment: 360-374
Protein B site: PxxP_1
Protein B sequence: EIKKIPPERPEMLPN
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0
Holdup BI standard deviation: 0.05
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.06.29.
Number of measurements: 2
The affinity was below the detection threshold of the assay.
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: SH3KBP1
Protein B fragment: 390-404
Protein B site: PxxP_2
Protein B sequence: LQKPSVPAIPPKKPR
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.36
Holdup BI standard deviation: 0.02
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: 4.52
Experiment series 8
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: SH3KBP1
Protein B fragment: 413-427
Protein B site: PxxP_3
Protein B sequence: RPGALPPRRPERPVG
Protein B construct: Biotinylated 15-mer peptide (crude)
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: 2022.06.29.
Number of measurements: 2
The affinity was below the detection threshold of the assay.
Experiment series 9
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: SH3KBP1
Protein B fragment: 360-374
Protein B site: PxxP_1
Protein B sequence: EIKKIPPERPEMLPN
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0
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
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 modification: D537V
Protein A construct: avi(biotin)-his6-MBP-TEVsite-SH3
Protein B protein: SH3KBP1
Protein B fragment: 390-404
Protein B site: PxxP_2
Protein B sequence: LQKPSVPAIPPKKPR
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.1
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: 3.78
Experiment series 11
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: SH3KBP1
Protein B fragment: 413-427
Protein B site: PxxP_3
Protein B sequence: RPGALPPRRPERPVG
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.01
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
The affinity was below the detection threshold of the assay.
Experiment series 12
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: SH3KBP1
Protein B fragment: 360-374
Protein B site: PxxP_1
Protein B sequence: EIKKIPPERPEMLPN
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 13
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: SH3KBP1
Protein B fragment: 390-404
Protein B site: PxxP_2
Protein B sequence: LQKPSVPAIPPKKPR
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0
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
The affinity was below the detection threshold of the assay.
Experiment series 14
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: SH3KBP1
Protein B fragment: 413-427
Protein B site: PxxP_3
Protein B sequence: RPGALPPRRPERPVG
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0
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 15
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: SH3KBP1
Protein B fragment: 360-374
Protein B site: PxxP_1
Protein B sequence: EIKKIPPERPEMLPN
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0
Holdup BI standard deviation: 0.05
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 16
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: SH3KBP1
Protein B fragment: 390-404
Protein B site: PxxP_2
Protein B sequence: LQKPSVPAIPPKKPR
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.14
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: 4.45
Experiment series 17
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: SH3KBP1
Protein B fragment: 413-427
Protein B site: PxxP_3
Protein B sequence: RPGALPPRRPERPVG
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: -0.02
Holdup BI standard deviation: 0.08
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 18
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: SH3KBP1
Protein B fragment: 360-374
Protein B site: PxxP_1
Protein B sequence: EIKKIPPERPEMLPN
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.03
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
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: Q540H
Protein A construct: avi(biotin)-his6-MBP-TEVsite-SH3
Protein B protein: SH3KBP1
Protein B fragment: 390-404
Protein B site: PxxP_2
Protein B sequence: LQKPSVPAIPPKKPR
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.33
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: 4.48
Experiment series 20
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: SH3KBP1
Protein B fragment: 413-427
Protein B site: PxxP_3
Protein B sequence: RPGALPPRRPERPVG
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.06
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: 3.54
Experiment series 21
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: SH3KBP1
Protein B fragment: 360-374
Protein B site: PxxP_1
Protein B sequence: EIKKIPPERPEMLPN
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: -0
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
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: T532M
Protein A construct: avi(biotin)-his6-MBP-TEVsite-SH3
Protein B protein: SH3KBP1
Protein B fragment: 390-404
Protein B site: PxxP_2
Protein B sequence: LQKPSVPAIPPKKPR
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.3
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: 4.4
Experiment series 23
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: SH3KBP1
Protein B fragment: 413-427
Protein B site: PxxP_3
Protein B sequence: RPGALPPRRPERPVG
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 24
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: SH3KBP1
Protein B fragment: 360-374
Protein B site: PxxP_1
Protein B sequence: EIKKIPPERPEMLPN
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 25
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: SH3KBP1
Protein B fragment: 390-404
Protein B site: PxxP_2
Protein B sequence: LQKPSVPAIPPKKPR
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.32
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: 4.45
Experiment series 26
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: SH3KBP1
Protein B fragment: 413-427
Protein B site: PxxP_3
Protein B sequence: RPGALPPRRPERPVG
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): 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 27
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: SH3KBP1
Protein B fragment: 360-374
Protein B site: PxxP_1
Protein B sequence: EIKKIPPERPEMLPN
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.02
Holdup BI standard deviation: 0.01
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 28
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: SH3KBP1
Protein B fragment: 390-404
Protein B site: PxxP_2
Protein B sequence: LQKPSVPAIPPKKPR
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.24
Holdup BI standard deviation: 0.03
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: 4.58
Experiment series 29
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: SH3KBP1
Protein B fragment: 413-427
Protein B site: PxxP_3
Protein B sequence: RPGALPPRRPERPVG
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.02
Holdup BI standard deviation: 0.01
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 30
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: SH3KBP1
Protein B fragment: 360-374
Protein B site: PxxP_1
Protein B sequence: EIKKIPPERPEMLPN
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.01
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 31
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: SH3KBP1
Protein B fragment: 390-404
Protein B site: PxxP_2
Protein B sequence: LQKPSVPAIPPKKPR
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.31
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: 4.42
Experiment series 32
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: SH3KBP1
Protein B fragment: 413-427
Protein B site: PxxP_3
Protein B sequence: RPGALPPRRPERPVG
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.06
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: 3.59
Experiment series 33
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: SH3KBP1
Protein B fragment: 360-374
Protein B site: PxxP_1
Protein B sequence: EIKKIPPERPEMLPN
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0
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
The affinity was below the detection threshold of the assay.
Experiment series 34
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: SH3KBP1
Protein B fragment: 390-404
Protein B site: PxxP_2
Protein B sequence: LQKPSVPAIPPKKPR
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.05
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: 3.5
Experiment series 35
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: SH3KBP1
Protein B fragment: 413-427
Protein B site: PxxP_3
Protein B sequence: RPGALPPRRPERPVG
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.02
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
The affinity was below the detection threshold of the assay.
Experiment series 36
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: SH3KBP1
Protein B fragment: 360-374
Protein B site: PxxP_1
Protein B sequence: EIKKIPPERPEMLPN
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): 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 37
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: SH3KBP1
Protein B fragment: 390-404
Protein B site: PxxP_2
Protein B sequence: LQKPSVPAIPPKKPR
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.14
Holdup BI standard deviation: 0.03
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: 4.47
Experiment series 38
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: SH3KBP1
Protein B fragment: 413-427
Protein B site: PxxP_3
Protein B sequence: RPGALPPRRPERPVG
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 39
Protein A protein: BIN1
Protein A fragment: 513-593
Protein A site: SH3
Protein A construct: his6-MBP-TEVsite-SH3
Protein B protein: SH3KBP1
Protein B fragment: 390-404
Protein B site: PxxP_2
Protein B sequence: LQKPSVPAIPPKKPR
Protein B construct: Bio E E Ado Ado-LQKPSVPAIPPKKPR-CONH2
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 40
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: SH3KBP1
Protein B fragment: 390-404
Protein B site: PxxP_2
Protein B sequence: LQKPSVPAIPPKKPR
Protein B construct: Bio E E Ado Ado-LQKPSVPAIPPKKPR-CONH2
Average holdup BI: 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: 1
The affinity was below the detection threshold of the assay.
Experiment series 41
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: SH3KBP1
Protein B fragment: 390-404
Protein B site: PxxP_2
Protein B sequence: LQKPSVPAIPPKKPR
Protein B construct: Bio E E Ado Ado-LQKPSVPAIPPKKPR-CONH2
Average holdup BI: 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: 2023/01/17
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 42
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: SH3KBP1
Protein B fragment: 390-404
Protein B site: PxxP_2
Protein B sequence: LQKPSVPAIPPKKPR
Protein B construct: Bio E E Ado Ado-LQKPSVPAIPPKKPR-CONH2
Average holdup BI: -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: 2023/01/17
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 43
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: SH3KBP1
Protein B fragment: 390-404
Protein B site: PxxP_2
Protein B sequence: LQKPSVPAIPPKKPR
Protein B construct: Bio E E Ado Ado-LQKPSVPAIPPKKPR-CONH2
Average holdup BI: 0.02
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 44
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: SH3KBP1
Protein B fragment: 390-404
Protein B site: PxxP_2
Protein B sequence: LQKPSVPAIPPKKPR
Protein B construct: Bio E E Ado Ado-LQKPSVPAIPPKKPR-CONH2
Average holdup BI: 0
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 45
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: SH3KBP1
Protein B fragment: 390-404
Protein B site: PxxP_2
Protein B sequence: LQKPSVPAIPPKKPR
Protein B construct: Bio E E Ado Ado-LQKPSVPAIPPKKPR-CONH2
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 46
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: SH3KBP1
Protein B fragment: 390-404
Protein B site: PxxP_2
Protein B sequence: LQKPSVPAIPPKKPR
Protein B construct: Bio E E Ado Ado-LQKPSVPAIPPKKPR-CONH2
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: 2023/01/18
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 47
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: SH3KBP1
Protein B fragment: 390-404
Protein B site: PxxP_2
Protein B sequence: LQKPSVPAIPPKKPR
Protein B construct: Bio E E Ado Ado-LQKPSVPAIPPKKPR-CONH2
Average holdup BI: 0.01
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 48
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: SH3KBP1
Protein B fragment: 390-404
Protein B site: PxxP_2
Protein B sequence: LQKPSVPAIPPKKPR
Protein B construct: Bio E E Ado Ado-LQKPSVPAIPPKKPR-CONH2
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: 2023/01/18
Number of measurements: 1
The affinity was below the detection threshold of the assay.