Details of experiments between O00499 and Q96J02
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: ITCH
Protein B fragment: 1-903
Protein B construct: endogenous protein (Jurkat)
Average holdup BI: 0.92
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): 8.66
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: 6.05
Experiment series 2
Protein A protein: BIN1
Protein A fragment: 513-593
Protein A site: SH3
Protein A construct: his6-MBP-TEVsite-SH3
Protein B protein: ITCH
Protein B fragment: 244-258
Protein B site: PxxP_1
Protein B sequence: SNGGFKPSRPPRPSR
Protein B construct: Bio Ado Ado-SNGGFKPSRPPRPSR-CONH2
Average holdup BI: 0.77
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: 5.35
Experiment series 3
Protein A protein: BIN1
Protein A fragment: 513-593
Protein A site: SH3
Protein A construct: his6-MBP-TEVsite-SH3
Protein B protein: ITCH
Protein B fragment: 247-261
Protein B site: PxxP_2
Protein B sequence: GFKPSRPPRPSRPPP
Protein B construct: Bio Ado Ado-GFKPSRPPRPSRPPP-CONH2
Average holdup BI: 0.71
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: 5.22
Experiment series 4
Protein A protein: BIN1
Protein A fragment: 513-593
Protein A site: SH3
Protein A construct: his6-MBP-TEVsite-SH3
Protein B protein: ITCH
Protein B fragment: 255-269
Protein B site: PxxP_3
Protein B sequence: RPSRPPPPTPRRPAS
Protein B construct: Bio Ado Ado-RPSRPPPPTPRRPAS-CONH2
Average holdup BI: 0.25
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: 4.28
Experiment series 5
Protein A protein: BIN1
Protein A fragment: 513-593
Protein A site: SH3
Protein A construct: his6-MBP-TEVsite-SH3
Protein B protein: ITCH
Protein B fragment: 353-367
Protein B site: PxxP_4
Protein B sequence: TWDRPEPLPPGWERR
Protein B construct: Bio Ado Ado-TWDRPEPLPPGWERR-CONH2
Average holdup BI: -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: 2023/01/16
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 6
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: ITCH
Protein B fragment: 244-258
Protein B site: PxxP_1
Protein B sequence: SNGGFKPSRPPRPSR
Protein B construct: Bio Ado Ado-SNGGFKPSRPPRPSR-CONH2
Average holdup BI: 0.79
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: 5.41
Experiment series 7
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: ITCH
Protein B fragment: 247-261
Protein B site: PxxP_2
Protein B sequence: GFKPSRPPRPSRPPP
Protein B construct: Bio Ado Ado-GFKPSRPPRPSRPPP-CONH2
Average holdup BI: 0.73
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: 5.24
Experiment series 8
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: ITCH
Protein B fragment: 255-269
Protein B site: PxxP_3
Protein B sequence: RPSRPPPPTPRRPAS
Protein B construct: Bio Ado Ado-RPSRPPPPTPRRPAS-CONH2
Average holdup BI: 0.24
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: 4.28
Experiment series 9
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: ITCH
Protein B fragment: 353-367
Protein B site: PxxP_4
Protein B sequence: TWDRPEPLPPGWERR
Protein B construct: Bio Ado Ado-TWDRPEPLPPGWERR-CONH2
Average holdup BI: -0.04
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 10
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: ITCH
Protein B fragment: 244-258
Protein B site: PxxP_1
Protein B sequence: SNGGFKPSRPPRPSR
Protein B construct: Bio Ado Ado-SNGGFKPSRPPRPSR-CONH2
Average holdup BI: 0.75
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: 5.3
Experiment series 11
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: ITCH
Protein B fragment: 247-261
Protein B site: PxxP_2
Protein B sequence: GFKPSRPPRPSRPPP
Protein B construct: Bio Ado Ado-GFKPSRPPRPSRPPP-CONH2
Average holdup BI: 0.72
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: 5.21
Experiment series 12
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: ITCH
Protein B fragment: 255-269
Protein B site: PxxP_3
Protein B sequence: RPSRPPPPTPRRPAS
Protein B construct: Bio Ado Ado-RPSRPPPPTPRRPAS-CONH2
Average holdup BI: 0.3
Holdup BI standard deviation: 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: 2
Measured pKd: 4.4
Experiment series 13
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: ITCH
Protein B fragment: 353-367
Protein B site: PxxP_4
Protein B sequence: TWDRPEPLPPGWERR
Protein B construct: Bio Ado Ado-TWDRPEPLPPGWERR-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 14
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: ITCH
Protein B fragment: 244-258
Protein B site: PxxP_1
Protein B sequence: SNGGFKPSRPPRPSR
Protein B construct: Bio Ado Ado-SNGGFKPSRPPRPSR-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: 2023/01/17
Number of measurements: 1
Measured pKd: 4.32
Experiment series 15
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: ITCH
Protein B fragment: 247-261
Protein B site: PxxP_2
Protein B sequence: GFKPSRPPRPSRPPP
Protein B construct: Bio Ado Ado-GFKPSRPPRPSRPPP-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
Measured pKd: 4
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: ITCH
Protein B fragment: 255-269
Protein B site: PxxP_3
Protein B sequence: RPSRPPPPTPRRPAS
Protein B construct: Bio Ado Ado-RPSRPPPPTPRRPAS-CONH2
Average holdup BI: 0.04
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 17
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: ITCH
Protein B fragment: 353-367
Protein B site: PxxP_4
Protein B sequence: TWDRPEPLPPGWERR
Protein B construct: Bio Ado Ado-TWDRPEPLPPGWERR-CONH2
Average holdup BI: -0.06
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 18
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: ITCH
Protein B fragment: 244-258
Protein B site: PxxP_1
Protein B sequence: SNGGFKPSRPPRPSR
Protein B construct: Bio Ado Ado-SNGGFKPSRPPRPSR-CONH2
Average holdup BI: 0.58
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: 5.12
Experiment series 19
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: ITCH
Protein B fragment: 247-261
Protein B site: PxxP_2
Protein B sequence: GFKPSRPPRPSRPPP
Protein B construct: Bio Ado Ado-GFKPSRPPRPSRPPP-CONH2
Average holdup BI: 0.53
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: 5.01
Experiment series 20
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: ITCH
Protein B fragment: 255-269
Protein B site: PxxP_3
Protein B sequence: RPSRPPPPTPRRPAS
Protein B construct: Bio Ado Ado-RPSRPPPPTPRRPAS-CONH2
Average holdup BI: 0.18
Holdup BI standard deviation: 0
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: 4.24
Experiment series 21
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: ITCH
Protein B fragment: 353-367
Protein B site: PxxP_4
Protein B sequence: TWDRPEPLPPGWERR
Protein B construct: Bio Ado Ado-TWDRPEPLPPGWERR-CONH2
Average holdup BI: -0.1
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 22
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: ITCH
Protein B fragment: 244-258
Protein B site: PxxP_1
Protein B sequence: SNGGFKPSRPPRPSR
Protein B construct: Bio Ado Ado-SNGGFKPSRPPRPSR-CONH2
Average holdup BI: 0.56
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: 5.29
Experiment series 23
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: ITCH
Protein B fragment: 247-261
Protein B site: PxxP_2
Protein B sequence: GFKPSRPPRPSRPPP
Protein B construct: Bio Ado Ado-GFKPSRPPRPSRPPP-CONH2
Average holdup BI: 0.5
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: 5.17
Experiment series 24
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: ITCH
Protein B fragment: 255-269
Protein B site: PxxP_3
Protein B sequence: RPSRPPPPTPRRPAS
Protein B construct: Bio Ado Ado-RPSRPPPPTPRRPAS-CONH2
Average holdup BI: 0.1
Holdup BI standard deviation: 0.02
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: 4.17
Experiment series 25
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: ITCH
Protein B fragment: 353-367
Protein B site: PxxP_4
Protein B sequence: TWDRPEPLPPGWERR
Protein B construct: Bio Ado Ado-TWDRPEPLPPGWERR-CONH2
Average holdup BI: -0.02
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 26
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: ITCH
Protein B fragment: 244-258
Protein B site: PxxP_1
Protein B sequence: SNGGFKPSRPPRPSR
Protein B construct: Bio Ado Ado-SNGGFKPSRPPRPSR-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 27
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: ITCH
Protein B fragment: 247-261
Protein B site: PxxP_2
Protein B sequence: GFKPSRPPRPSRPPP
Protein B construct: Bio Ado Ado-GFKPSRPPRPSRPPP-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 28
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: ITCH
Protein B fragment: 255-269
Protein B site: PxxP_3
Protein B sequence: RPSRPPPPTPRRPAS
Protein B construct: Bio Ado Ado-RPSRPPPPTPRRPAS-CONH2
Average holdup BI: -0.03
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 29
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: ITCH
Protein B fragment: 353-367
Protein B site: PxxP_4
Protein B sequence: TWDRPEPLPPGWERR
Protein B construct: Bio Ado Ado-TWDRPEPLPPGWERR-CONH2
Average holdup BI: -0.06
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 30
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: ITCH
Protein B fragment: 244-258
Protein B site: PxxP_1
Protein B sequence: SNGGFKPSRPPRPSR
Protein B construct: Bio Ado Ado-SNGGFKPSRPPRPSR-CONH2
Average holdup BI: 0.69
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: 5.17
Experiment series 31
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: ITCH
Protein B fragment: 247-261
Protein B site: PxxP_2
Protein B sequence: GFKPSRPPRPSRPPP
Protein B construct: Bio Ado Ado-GFKPSRPPRPSRPPP-CONH2
Average holdup BI: 0.63
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: 5.04
Experiment series 32
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: ITCH
Protein B fragment: 255-269
Protein B site: PxxP_3
Protein B sequence: RPSRPPPPTPRRPAS
Protein B construct: Bio Ado Ado-RPSRPPPPTPRRPAS-CONH2
Average holdup BI: 0.03
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: 2023/01/18
Number of measurements: 2
The affinity was below the detection threshold of the assay.
Experiment series 33
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: ITCH
Protein B fragment: 353-367
Protein B site: PxxP_4
Protein B sequence: TWDRPEPLPPGWERR
Protein B construct: Bio Ado Ado-TWDRPEPLPPGWERR-CONH2
Average holdup BI: -0.07
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 34
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: ITCH
Protein B fragment: 244-258
Protein B site: PxxP_1
Protein B sequence: SNGGFKPSRPPRPSR
Protein B construct: Bio Ado Ado-SNGGFKPSRPPRPSR-CONH2
Average holdup BI: 0.61
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: 5.32
Experiment series 35
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: ITCH
Protein B fragment: 247-261
Protein B site: PxxP_2
Protein B sequence: GFKPSRPPRPSRPPP
Protein B construct: Bio Ado Ado-GFKPSRPPRPSRPPP-CONH2
Average holdup BI: 0.56
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: 5.2
Experiment series 36
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: ITCH
Protein B fragment: 255-269
Protein B site: PxxP_3
Protein B sequence: RPSRPPPPTPRRPAS
Protein B construct: Bio Ado Ado-RPSRPPPPTPRRPAS-CONH2
Average holdup BI: 0.13
Holdup BI standard deviation: 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: 2
Measured pKd: 4.19
Experiment series 37
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: ITCH
Protein B fragment: 353-367
Protein B site: PxxP_4
Protein B sequence: TWDRPEPLPPGWERR
Protein B construct: Bio Ado Ado-TWDRPEPLPPGWERR-CONH2
Average holdup BI: -0.09
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 38
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: ITCH
Protein B fragment: 244-258
Protein B site: PxxP_1
Protein B sequence: SNGGFKPSRPPRPSR
Protein B construct: Bio Ado Ado-SNGGFKPSRPPRPSR-CONH2
Average holdup BI: 0.74
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: 5.28
Experiment series 39
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: ITCH
Protein B fragment: 247-261
Protein B site: PxxP_2
Protein B sequence: GFKPSRPPRPSRPPP
Protein B construct: Bio Ado Ado-GFKPSRPPRPSRPPP-CONH2
Average holdup BI: 0.69
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: 5.15
Experiment series 40
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: ITCH
Protein B fragment: 255-269
Protein B site: PxxP_3
Protein B sequence: RPSRPPPPTPRRPAS
Protein B construct: Bio Ado Ado-RPSRPPPPTPRRPAS-CONH2
Average holdup BI: 0.27
Holdup BI standard deviation: 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: 2023/01/18
Number of measurements: 2
Measured pKd: 4.34
Experiment series 41
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: ITCH
Protein B fragment: 353-367
Protein B site: PxxP_4
Protein B sequence: TWDRPEPLPPGWERR
Protein B construct: Bio Ado Ado-TWDRPEPLPPGWERR-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.