Details of experiments between the fragments Q8IYB3~365-379 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: SRRM1
Protein A fragment: 365-379
Protein A site: PxxP_2
Protein A sequence: SRSRSPPKKPPKRTS
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: 0.51
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: 4
Measured pKd: 4.82
Experiment series 2
Protein A protein: SRRM1
Protein A fragment: 365-379
Protein A site: PxxP_2
Protein A sequence: SRSRSPPKKPPKRTS
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: 0.46
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: 4
Measured pKd: 4.72
Experiment series 3
Protein A protein: SRRM1
Protein A fragment: 365-379
Protein A site: PxxP_2
Protein A sequence: SRSRSPPKKPPKRTS
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: 0.15
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: 4
Measured pKd: 4.01
Experiment series 4
Protein A protein: SRRM1
Protein A fragment: 365-379
Protein A site: PxxP_2
Protein A sequence: SRSRSPPKKPPKRTS
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: -0.01
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: 4
The affinity was below the detection threshold of the assay.
Experiment series 5
Protein A protein: SRRM1
Protein A fragment: 365-379
Protein A site: PxxP_2
Protein A sequence: SRSRSPPKKPPKRTS
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: 0.21
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: 4
Measured pKd: 4.7
Experiment series 6
Protein A protein: SRRM1
Protein A fragment: 365-379
Protein A site: PxxP_2
Protein A sequence: SRSRSPPKKPPKRTS
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: 0.46
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: 4
Measured pKd: 4.72
Experiment series 7
Protein A protein: SRRM1
Protein A fragment: 365-379
Protein A site: PxxP_2
Protein A sequence: SRSRSPPKKPPKRTS
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: 0.4
Holdup BI standard deviation: 0.05
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: 4.62
Experiment series 8
Protein A protein: SRRM1
Protein A fragment: 365-379
Protein A site: PxxP_2
Protein A sequence: SRSRSPPKKPPKRTS
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: 0.42
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: 4
Measured pKd: 4.66
Experiment series 9
Protein A protein: SRRM1
Protein A fragment: 365-379
Protein A site: PxxP_2
Protein A sequence: SRSRSPPKKPPKRTS
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: 0.37
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: 4
Measured pKd: 4.9
Experiment series 10
Protein A protein: SRRM1
Protein A fragment: 365-379
Protein A site: PxxP_2
Protein A sequence: SRSRSPPKKPPKRTS
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: 0.02
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: 4.68
Experiment series 11
Protein A protein: SRRM1
Protein A fragment: 365-379
Protein A site: PxxP_2
Protein A sequence: SRSRSPPKKPPKRTS
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: 0.53
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: 4.84
Experiment series 12
Protein A protein: SRRM1
Protein A fragment: 365-379
Protein A site: PxxP_2
Protein A sequence: SRSRSPPKKPPKRTS
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: 0.22
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: 4
Measured pKd: 4.71