Details of experiments between the fragments O00499~513-593 and A6NHR9~1977-1991
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: SMCHD1
Protein B fragment: 1977-1991
Protein B site: PxxP_2
Protein B sequence: VETTDCPVPPKRMRR
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.88
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: 5.69
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: SMCHD1
Protein B fragment: 1977-1991
Protein B site: PxxP_2
Protein B sequence: VETTDCPVPPKRMRR
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.84
Holdup BI standard deviation: 0
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: 5.57
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: SMCHD1
Protein B fragment: 1977-1991
Protein B site: PxxP_2
Protein B sequence: VETTDCPVPPKRMRR
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.43
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: 4.67
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: SMCHD1
Protein B fragment: 1977-1991
Protein B site: PxxP_2
Protein B sequence: VETTDCPVPPKRMRR
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: -0.01
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
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: SMCHD1
Protein B fragment: 1977-1991
Protein B site: PxxP_2
Protein B sequence: VETTDCPVPPKRMRR
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.64
Holdup BI standard deviation: 0
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: 5.62
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: SMCHD1
Protein B fragment: 1977-1991
Protein B site: PxxP_2
Protein B sequence: VETTDCPVPPKRMRR
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.79
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: 5.4
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: SMCHD1
Protein B fragment: 1977-1991
Protein B site: PxxP_2
Protein B sequence: VETTDCPVPPKRMRR
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.81
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: 5.47
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: SMCHD1
Protein B fragment: 1977-1991
Protein B site: PxxP_2
Protein B sequence: VETTDCPVPPKRMRR
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.77
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: 5.34
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: SMCHD1
Protein B fragment: 1977-1991
Protein B site: PxxP_2
Protein B sequence: VETTDCPVPPKRMRR
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.71
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
Measured pKd: 5.61
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: SMCHD1
Protein B fragment: 1977-1991
Protein B site: PxxP_2
Protein B sequence: VETTDCPVPPKRMRR
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.83
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: 5.51
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: SMCHD1
Protein B fragment: 1977-1991
Protein B site: PxxP_2
Protein B sequence: VETTDCPVPPKRMRR
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.45
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: 4.71
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: SMCHD1
Protein B fragment: 1977-1991
Protein B site: PxxP_2
Protein B sequence: VETTDCPVPPKRMRR
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.68
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
Measured pKd: 5.7
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: SMCHD1
Protein B fragment: 1977-1991
Protein B site: PxxP_2
Protein B sequence: VETTDCPVPPKRMRR
Protein B construct: Bio E E Ado Ado-VETTDCPVPPKRMRR-CONH2
Average holdup BI: 0.47
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
Measured pKd: 4.74
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: SMCHD1
Protein B fragment: 1977-1991
Protein B site: PxxP_2
Protein B sequence: VETTDCPVPPKRMRR
Protein B construct: Bio E E Ado Ado-VETTDCPVPPKRMRR-CONH2
Average holdup BI: 0.48
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
Measured pKd: 4.76
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: SMCHD1
Protein B fragment: 1977-1991
Protein B site: PxxP_2
Protein B sequence: VETTDCPVPPKRMRR
Protein B construct: Bio E E Ado Ado-VETTDCPVPPKRMRR-CONH2
Average holdup BI: 0.52
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
Measured pKd: 4.82
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: SMCHD1
Protein B fragment: 1977-1991
Protein B site: PxxP_2
Protein B sequence: VETTDCPVPPKRMRR
Protein B construct: Bio E E Ado Ado-VETTDCPVPPKRMRR-CONH2
Average holdup BI: 0.11
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
Measured pKd: 3.87
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: SMCHD1
Protein B fragment: 1977-1991
Protein B site: PxxP_2
Protein B sequence: VETTDCPVPPKRMRR
Protein B construct: Bio E E Ado Ado-VETTDCPVPPKRMRR-CONH2
Average holdup BI: 0.37
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
Measured pKd: 4.7
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: SMCHD1
Protein B fragment: 1977-1991
Protein B site: PxxP_2
Protein B sequence: VETTDCPVPPKRMRR
Protein B construct: Bio E E Ado Ado-VETTDCPVPPKRMRR-CONH2
Average holdup BI: 0.31
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
Measured pKd: 4.79
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: SMCHD1
Protein B fragment: 1977-1991
Protein B site: PxxP_2
Protein B sequence: VETTDCPVPPKRMRR
Protein B construct: Bio E E Ado Ado-VETTDCPVPPKRMRR-CONH2
Average holdup BI: -0.15
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: BIN1
Protein A fragment: 513-593
Protein A site: SH3
Protein A modification: Y531S
Protein A construct: his6-MBP-TEVsite-SH3
Protein B protein: SMCHD1
Protein B fragment: 1977-1991
Protein B site: PxxP_2
Protein B sequence: VETTDCPVPPKRMRR
Protein B construct: Bio E E Ado Ado-VETTDCPVPPKRMRR-CONH2
Average holdup BI: 0.14
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
Measured pKd: 3.96
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: SMCHD1
Protein B fragment: 1977-1991
Protein B site: PxxP_2
Protein B sequence: VETTDCPVPPKRMRR
Protein B construct: Bio E E Ado Ado-VETTDCPVPPKRMRR-CONH2
Average holdup BI: 0.31
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
Measured pKd: 4.72
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: SMCHD1
Protein B fragment: 1977-1991
Protein B site: PxxP_2
Protein B sequence: VETTDCPVPPKRMRR
Protein B construct: Bio E E Ado Ado-VETTDCPVPPKRMRR-CONH2
Average holdup BI: 0.53
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
Measured pKd: 4.85