Details of experiments between the fragments P38159~90-104 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: RBMX
Protein A fragment: 90-104
Protein A site: PxxP_2
Protein A sequence: ESGRRGPPPPPRSRG
Protein A construct: Bio Ado Ado-ESGRRGPPPPPRSRG-CONH2
Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.44
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: 4.69
Experiment series 2
Protein A protein: RBMX
Protein A fragment: 90-104
Protein A site: PxxP_2
Protein A sequence: ESGRRGPPPPPRSRG
Protein A construct: Bio Ado Ado-ESGRRGPPPPPRSRG-CONH2
Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: T532M
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.46
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: 4.72
Experiment series 3
Protein A protein: RBMX
Protein A fragment: 90-104
Protein A site: PxxP_2
Protein A sequence: ESGRRGPPPPPRSRG
Protein A construct: Bio Ado Ado-ESGRRGPPPPPRSRG-CONH2
Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: Q540H
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.46
Holdup BI standard deviation: 0.08
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: 4.71
Experiment series 4
Protein A protein: RBMX
Protein A fragment: 90-104
Protein A site: PxxP_2
Protein A sequence: ESGRRGPPPPPRSRG
Protein A construct: Bio Ado Ado-ESGRRGPPPPPRSRG-CONH2
Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: D537V
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.05
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: 2023/01/17
Number of measurements: 2
Measured pKd: 3.49
Experiment series 5
Protein A protein: RBMX
Protein A fragment: 90-104
Protein A site: PxxP_2
Protein A sequence: ESGRRGPPPPPRSRG
Protein A construct: Bio Ado Ado-ESGRRGPPPPPRSRG-CONH2
Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: V583I
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.28
Holdup BI standard deviation: 0.01
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
Measured pKd: 4.52
Experiment series 6
Protein A protein: RBMX
Protein A fragment: 90-104
Protein A site: PxxP_2
Protein A sequence: ESGRRGPPPPPRSRG
Protein A construct: Bio Ado Ado-ESGRRGPPPPPRSRG-CONH2
Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: P551L
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.23
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
Measured pKd: 4.61
Experiment series 7
Protein A protein: RBMX
Protein A fragment: 90-104
Protein A site: PxxP_2
Protein A sequence: ESGRRGPPPPPRSRG
Protein A construct: Bio Ado Ado-ESGRRGPPPPPRSRG-CONH2
Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: F584S
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: -0.01
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: 2023/01/17
Number of measurements: 2
The affinity was below the detection threshold of the assay.
Experiment series 8
Protein A protein: RBMX
Protein A fragment: 90-104
Protein A site: PxxP_2
Protein A sequence: ESGRRGPPPPPRSRG
Protein A construct: Bio Ado Ado-ESGRRGPPPPPRSRG-CONH2
Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: Y531S
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.07
Holdup BI standard deviation: 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: 2
Measured pKd: 3.61
Experiment series 9
Protein A protein: RBMX
Protein A fragment: 90-104
Protein A site: PxxP_2
Protein A sequence: ESGRRGPPPPPRSRG
Protein A construct: Bio Ado Ado-ESGRRGPPPPPRSRG-CONH2
Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: R581C
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.26
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
Measured pKd: 4.59
Experiment series 10
Protein A protein: RBMX
Protein A fragment: 90-104
Protein A site: PxxP_2
Protein A sequence: ESGRRGPPPPPRSRG
Protein A construct: Bio Ado Ado-ESGRRGPPPPPRSRG-CONH2
Protein B protein: BIN1
Protein B fragment: 513-593
Protein B site: SH3
Protein B modification: V566M
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.47
Holdup BI standard deviation: 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: 2
Measured pKd: 4.73