Details of experiments between O00499 and Q29RF7
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: PDS5A
Protein B fragment: 1-1337
Protein B construct: endogenous protein (Jurkat)
Average holdup BI: 0.37
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): 6.51
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: 4.77
Experiment series 2
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: PDS5A
Protein B fragment: 1-1337
Protein B construct: endogenous protein (Jurkat)
Average holdup BI: 0.26
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): 3.5
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
The affinity was below the detection threshold of the assay.
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: PDS5A
Protein B fragment: 1256-1270
Protein B site: PxxP_1
Protein B sequence: KVDESGPPAPSKPRR
Protein B construct: Bio Ado Ado-KVDESGPPAPSKPRR-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: 2023/01/16
Number of measurements: 1
Measured pKd: 3.84
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: PDS5A
Protein B fragment: 1259-1273
Protein B site: PxxP_2
Protein B sequence: ESGPPAPSKPRRGRR
Protein B construct: Bio Ado Ado-ESGPPAPSKPRRGRR-CONH2
Average holdup BI: 0.51
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: 4.82
Experiment series 5
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: PDS5A
Protein B fragment: 1256-1270
Protein B site: PxxP_1
Protein B sequence: KVDESGPPAPSKPRR
Protein B construct: Bio Ado Ado-KVDESGPPAPSKPRR-CONH2
Average holdup BI: 0.12
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: 3.9
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: PDS5A
Protein B fragment: 1259-1273
Protein B site: PxxP_2
Protein B sequence: ESGPPAPSKPRRGRR
Protein B construct: Bio Ado Ado-ESGPPAPSKPRRGRR-CONH2
Average holdup BI: 0.54
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: 4.87
Experiment series 7
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: PDS5A
Protein B fragment: 1256-1270
Protein B site: PxxP_1
Protein B sequence: KVDESGPPAPSKPRR
Protein B construct: Bio Ado Ado-KVDESGPPAPSKPRR-CONH2
Average holdup BI: 0.22
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: 4.21
Experiment series 8
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: PDS5A
Protein B fragment: 1259-1273
Protein B site: PxxP_2
Protein B sequence: ESGPPAPSKPRRGRR
Protein B construct: Bio Ado Ado-ESGPPAPSKPRRGRR-CONH2
Average holdup BI: 0.6
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: 4.98
Experiment series 9
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: PDS5A
Protein B fragment: 1256-1270
Protein B site: PxxP_1
Protein B sequence: KVDESGPPAPSKPRR
Protein B construct: Bio Ado Ado-KVDESGPPAPSKPRR-CONH2
Average holdup BI: 0
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023/01/17
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: D537V
Protein A construct: his6-MBP-TEVsite-SH3
Protein B protein: PDS5A
Protein B fragment: 1259-1273
Protein B site: PxxP_2
Protein B sequence: ESGPPAPSKPRRGRR
Protein B construct: Bio Ado Ado-ESGPPAPSKPRRGRR-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: 2023/01/17
Number of measurements: 1
Measured pKd: 3.69
Experiment series 11
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: PDS5A
Protein B fragment: 1256-1270
Protein B site: PxxP_1
Protein B sequence: KVDESGPPAPSKPRR
Protein B construct: Bio Ado Ado-KVDESGPPAPSKPRR-CONH2
Average holdup BI: 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: 1
The affinity was below the detection threshold of the assay.
Experiment series 12
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: PDS5A
Protein B fragment: 1259-1273
Protein B site: PxxP_2
Protein B sequence: ESGPPAPSKPRRGRR
Protein B construct: Bio Ado Ado-ESGPPAPSKPRRGRR-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: 4.71
Experiment series 13
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: PDS5A
Protein B fragment: 1256-1270
Protein B site: PxxP_1
Protein B sequence: KVDESGPPAPSKPRR
Protein B construct: Bio Ado Ado-KVDESGPPAPSKPRR-CONH2
Average holdup BI: 0.03
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 14
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: PDS5A
Protein B fragment: 1259-1273
Protein B site: PxxP_2
Protein B sequence: ESGPPAPSKPRRGRR
Protein B construct: Bio Ado Ado-ESGPPAPSKPRRGRR-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: 4.8
Experiment series 15
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: PDS5A
Protein B fragment: 1256-1270
Protein B site: PxxP_1
Protein B sequence: KVDESGPPAPSKPRR
Protein B construct: Bio Ado Ado-KVDESGPPAPSKPRR-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 16
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: PDS5A
Protein B fragment: 1259-1273
Protein B site: PxxP_2
Protein B sequence: ESGPPAPSKPRRGRR
Protein B construct: Bio Ado Ado-ESGPPAPSKPRRGRR-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 17
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: PDS5A
Protein B fragment: 1256-1270
Protein B site: PxxP_1
Protein B sequence: KVDESGPPAPSKPRR
Protein B construct: Bio Ado Ado-KVDESGPPAPSKPRR-CONH2
Average holdup BI: 0.27
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: 4.34
Experiment series 18
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: PDS5A
Protein B fragment: 1259-1273
Protein B site: PxxP_2
Protein B sequence: ESGPPAPSKPRRGRR
Protein B construct: Bio Ado Ado-ESGPPAPSKPRRGRR-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 19
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: PDS5A
Protein B fragment: 1256-1270
Protein B site: PxxP_1
Protein B sequence: KVDESGPPAPSKPRR
Protein B construct: Bio Ado Ado-KVDESGPPAPSKPRR-CONH2
Average holdup BI: 0.04
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 20
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: PDS5A
Protein B fragment: 1259-1273
Protein B site: PxxP_2
Protein B sequence: ESGPPAPSKPRRGRR
Protein B construct: Bio Ado Ado-ESGPPAPSKPRRGRR-CONH2
Average holdup BI: 0.34
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: 4.77
Experiment series 21
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: PDS5A
Protein B fragment: 1256-1270
Protein B site: PxxP_1
Protein B sequence: KVDESGPPAPSKPRR
Protein B construct: Bio Ado Ado-KVDESGPPAPSKPRR-CONH2
Average holdup BI: 0.12
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: 3.87
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: PDS5A
Protein B fragment: 1259-1273
Protein B site: PxxP_2
Protein B sequence: ESGPPAPSKPRRGRR
Protein B construct: Bio Ado Ado-ESGPPAPSKPRRGRR-CONH2
Average holdup BI: 0.55
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: 4.89