Details of experiments between the fragments O00499~513-593 and Q8IYB3~576-590
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: SRRM1
Protein B fragment: 576-590
Protein B site: PxxP_7
Protein B sequence: PPRRRTPSPPPRRRS
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.9
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: 4
Measured pKd: 5.8
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: SRRM1
Protein B fragment: 576-590
Protein B site: PxxP_7
Protein B sequence: PPRRRTPSPPPRRRS
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.88
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: 4
Measured pKd: 5.72
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: SRRM1
Protein B fragment: 576-590
Protein B site: PxxP_7
Protein B sequence: PPRRRTPSPPPRRRS
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.69
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: 4
Measured pKd: 5.16
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: SRRM1
Protein B fragment: 576-590
Protein B site: PxxP_7
Protein B sequence: PPRRRTPSPPPRRRS
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.12
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: 4
Measured pKd: 3.88
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: SRRM1
Protein B fragment: 576-590
Protein B site: PxxP_7
Protein B sequence: PPRRRTPSPPPRRRS
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.69
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: 4
Measured pKd: 5.73
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: SRRM1
Protein B fragment: 576-590
Protein B site: PxxP_7
Protein B sequence: PPRRRTPSPPPRRRS
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.83
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: 4
Measured pKd: 5.5
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: SRRM1
Protein B fragment: 576-590
Protein B site: PxxP_7
Protein B sequence: PPRRRTPSPPPRRRS
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.87
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: 4
Measured pKd: 5.66
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: SRRM1
Protein B fragment: 576-590
Protein B site: PxxP_7
Protein B sequence: PPRRRTPSPPPRRRS
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.81
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: 4
Measured pKd: 5.47
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: SRRM1
Protein B fragment: 576-590
Protein B site: PxxP_7
Protein B sequence: PPRRRTPSPPPRRRS
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): 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: 4
Measured pKd: 5.88
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: SRRM1
Protein B fragment: 576-590
Protein B site: PxxP_7
Protein B sequence: PPRRRTPSPPPRRRS
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.87
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: 4
Measured pKd: 5.67
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: SRRM1
Protein B fragment: 576-590
Protein B site: PxxP_7
Protein B sequence: PPRRRTPSPPPRRRS
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.64
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: 4
Measured pKd: 5.06
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: SRRM1
Protein B fragment: 576-590
Protein B site: PxxP_7
Protein B sequence: PPRRRTPSPPPRRRS
Protein B construct: Biotinylated 15-mer peptide (crude)
Average holdup BI: 0.75
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: 4
Measured pKd: 5.87
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: SRRM1
Protein B fragment: 576-590
Protein B site: PxxP_7
Protein B sequence: PPRRRTPSPPPRRRS
Protein B construct: Bio E E Ado Ado-PPRRRTPSPPPRRRS-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: 2023/01/16
Number of measurements: 1
Measured pKd: 4.86
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: SRRM1
Protein B fragment: 576-590
Protein B site: PxxP_7
Protein B sequence: PPRRRTPSPPPRRRS
Protein B construct: Bio E E Ado Ado-PPRRRTPSPPPRRRS-CONH2
Average holdup BI: 0.58
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: 4.94
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: SRRM1
Protein B fragment: 576-590
Protein B site: PxxP_7
Protein B sequence: PPRRRTPSPPPRRRS
Protein B construct: Bio E E Ado Ado-PPRRRTPSPPPRRRS-CONH2
Average holdup BI: 0.64
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: 5.05
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: SRRM1
Protein B fragment: 576-590
Protein B site: PxxP_7
Protein B sequence: PPRRRTPSPPPRRRS
Protein B construct: Bio E E Ado Ado-PPRRRTPSPPPRRRS-CONH2
Average holdup BI: 0.35
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: 4.51
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: SRRM1
Protein B fragment: 576-590
Protein B site: PxxP_7
Protein B sequence: PPRRRTPSPPPRRRS
Protein B construct: Bio E E Ado Ado-PPRRRTPSPPPRRRS-CONH2
Average holdup BI: 0.47
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: 4.9
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: SRRM1
Protein B fragment: 576-590
Protein B site: PxxP_7
Protein B sequence: PPRRRTPSPPPRRRS
Protein B construct: Bio E E Ado Ado-PPRRRTPSPPPRRRS-CONH2
Average holdup BI: 0.39
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: 4.96
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: SRRM1
Protein B fragment: 576-590
Protein B site: PxxP_7
Protein B sequence: PPRRRTPSPPPRRRS
Protein B construct: Bio E E Ado Ado-PPRRRTPSPPPRRRS-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 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: SRRM1
Protein B fragment: 576-590
Protein B site: PxxP_7
Protein B sequence: PPRRRTPSPPPRRRS
Protein B construct: Bio E E Ado Ado-PPRRRTPSPPPRRRS-CONH2
Average holdup BI: 0.26
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: 4.33
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: SRRM1
Protein B fragment: 576-590
Protein B site: PxxP_7
Protein B sequence: PPRRRTPSPPPRRRS
Protein B construct: Bio E E Ado Ado-PPRRRTPSPPPRRRS-CONH2
Average holdup BI: 0.37
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: 4.83
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: SRRM1
Protein B fragment: 576-590
Protein B site: PxxP_7
Protein B sequence: PPRRRTPSPPPRRRS
Protein B construct: Bio E E Ado Ado-PPRRRTPSPPPRRRS-CONH2
Average holdup BI: 0.61
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: 5