Details of experiments between P22681 and P00519
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: CBL
Protein A fragment: 1-906
Protein A construct: endogenous protein (Jurkat)
Protein B protein: ABL1
Protein B fragment: 56-121
Protein B site: SH3
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: 0.08
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): 0.56
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 2
Protein A protein: CBL
Protein A fragment: 488-502
Protein A site: PxxP_1
Protein A sequence: APQASLPPVPPRLDL
Protein A construct: Biotinylated 15-mer peptide (crude)
Protein B protein: ABL1
Protein B fragment: 56-121
Protein B site: SH3
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: 0.06
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: 2
Measured pKd: 3.55
Experiment series 3
Protein A protein: CBL
Protein A fragment: 528-542
Protein A site: PxxP_2
Protein A sequence: HKDKPLPVPPTLRDL
Protein A construct: Biotinylated 15-mer peptide (crude)
Protein B protein: ABL1
Protein B fragment: 56-121
Protein B site: SH3
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: 0
Holdup BI standard deviation: 0.01
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.06.29.
Number of measurements: 2
The affinity was below the detection threshold of the assay.
Experiment series 4
Protein A protein: CBL
Protein A fragment: 537-551
Protein A site: PxxP_3
Protein A sequence: PTLRDLPPPPPPDRP
Protein A construct: Biotinylated 15-mer peptide (crude)
Protein B protein: ABL1
Protein B fragment: 56-121
Protein B site: SH3
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: 0.07
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: 2
Measured pKd: 3.64
Experiment series 5
Protein A protein: CBL
Protein A fragment: 539-553
Protein A site: PxxP_4
Protein A sequence: LRDLPPPPPPDRPYS
Protein A construct: Biotinylated 15-mer peptide (crude)
Protein B protein: ABL1
Protein B fragment: 56-121
Protein B site: SH3
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: 0.09
Holdup BI standard deviation: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.06.29.
Number of measurements: 2
Measured pKd: 3.77
Experiment series 6
Protein A protein: CBL
Protein A fragment: 560-574
Protein A site: PxxP_5
Protein A sequence: PQRRPLPCTPGDCPS
Protein A construct: Biotinylated 15-mer peptide (crude)
Protein B protein: ABL1
Protein B fragment: 56-121
Protein B site: SH3
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: 0.01
Holdup BI standard deviation: 0.03
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.06.29.
Number of measurements: 2
The affinity was below the detection threshold of the assay.
Experiment series 7
Protein A protein: CBL
Protein A fragment: 590-604
Protein A site: PxxP_6
Protein A sequence: WLPRPIPKVPVSAPS
Protein A construct: Biotinylated 15-mer peptide (crude)
Protein B protein: ABL1
Protein B fragment: 56-121
Protein B site: SH3
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: 0.03
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: 2
The affinity was below the detection threshold of the assay.
Experiment series 8
Protein A protein: CBL
Protein A fragment: 814-828
Protein A site: PxxP_7
Protein A sequence: TEGSQVPERPPKPFP
Protein A construct: Biotinylated 15-mer peptide (crude)
Protein B protein: ABL1
Protein B fragment: 56-121
Protein B site: SH3
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: 0
Holdup BI standard deviation: 0.01
Immobilized partner concentration in holdup experiment (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022.06.29.
Number of measurements: 2
The affinity was below the detection threshold of the assay.