Details of experiments between P42768 and P55345
Note that the results of all experiments are listed, regardless of the modification states of the fragments.
Experiment series 1
Protein A protein: WAS
Protein A fragment: 1-502
Protein A construct: endogenous protein (Jurkat)
Protein B protein: PRMT2
Protein B fragment: 24-96
Protein B site: SH3
Protein B construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: 0.98
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): 7.28
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: 6.69
Experiment series 2
Protein A protein: WAS
Protein A fragment: 154-168
Protein A site: PxxP_1
Protein A sequence: GDRRQLPPPPTPANE
Protein A construct: Bio Ado Ado-GDRRQLPPPPTPANE-CONH2
Protein B protein: PRMT2
Protein B fragment: 24-96
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: -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: 2023/01/16
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 3
Protein A protein: WAS
Protein A fragment: 306-320
Protein A site: PxxP_2
Protein A sequence: EMRRQEPLPPPPPPS
Protein A construct: Bio Ado Ado-EMRRQEPLPPPPPPS-CONH2
Protein B protein: PRMT2
Protein B fragment: 24-96
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3
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: 2023/01/16
Number of measurements: 1
Measured pKd: 3.57
Experiment series 4
Protein A protein: WAS
Protein A fragment: 310-324
Protein A site: PxxP_3
Protein A sequence: QEPLPPPPPPSRGGN
Protein A construct: Bio Ado Ado-QEPLPPPPPPSRGGN-CONH2
Protein B protein: PRMT2
Protein B fragment: 24-96
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.03
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 5
Protein A protein: WAS
Protein A fragment: 335-349
Protein A site: PxxP_4
Protein A sequence: NKGRSGPLPPVPLGI
Protein A construct: Bio Ado Ado-NKGRSGPLPPVPLGI-CONH2
Protein B protein: PRMT2
Protein B fragment: 24-96
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3
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: 2023/01/16
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 6
Protein A protein: WAS
Protein A fragment: 353-367
Protein A site: PxxP_5
Protein A sequence: PPTPRGPPPPGRGGP
Protein A construct: Bio Ado Ado-PPTPRGPPPPGRGGP-CONH2
Protein B protein: PRMT2
Protein B fragment: 24-96
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3
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: 2023/01/16
Number of measurements: 1
The affinity was below the detection threshold of the assay.
Experiment series 7
Protein A protein: WAS
Protein A fragment: 361-375
Protein A site: PxxP_6
Protein A sequence: PPGRGGPPPPPPPAT
Protein A construct: Bio Ado Ado-PPGRGGPPPPPPPAT-CONH2
Protein B protein: PRMT2
Protein B fragment: 24-96
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3
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 8
Protein A protein: WAS
Protein A fragment: 374-388
Protein A site: PxxP_7
Protein A sequence: ATGRSGPLPPPPPGA
Protein A construct: Bio Ado Ado-ATGRSGPLPPPPPGA-CONH2
Protein B protein: PRMT2
Protein B fragment: 24-96
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3
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: 2023/01/16
Number of measurements: 1
The affinity was below the detection threshold of the assay.