Details of experiments between Q8IZ21 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: PHACTR4
Protein A fragment: 1-702
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: 0.09
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): 0.46
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: PHACTR4
Protein A fragment: 161-175
Protein A site: PxxP_1
Protein A sequence: ENVPKPPLLPPKRPL
Protein A construct: Bio Ado Ado-ENVPKPPLLPPKRPL-CONH2
Protein B protein: PRMT2
Protein B fragment: 24-96
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.23
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: 4.24
Experiment series 3
Protein A protein: PHACTR4
Protein A fragment: 229-243
Protein A site: PxxP_2
Protein A sequence: PAPRTLPAAPASTNT
Protein A construct: Bio Ado Ado-PAPRTLPAAPASTNT-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 4
Protein A protein: PHACTR4
Protein A fragment: 253-267
Protein A site: PxxP_3
Protein A sequence: VPAKQPPIPPPKPAH
Protein A construct: Bio Ado Ado-VPAKQPPIPPPKPAH-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
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
The affinity was below the detection threshold of the assay.
Experiment series 5
Protein A protein: PHACTR4
Protein A fragment: 286-300
Protein A site: PxxP_4
Protein A sequence: LLSKPSPPLPPKRGI
Protein A construct: Bio Ado Ado-LLSKPSPPLPPKRGI-CONH2
Protein B protein: PRMT2
Protein B fragment: 24-96
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: -0.07
Holdup BI standard deviation: 0.09
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
The affinity was below the detection threshold of the assay.
Experiment series 6
Protein A protein: PHACTR4
Protein A fragment: 287-301
Protein A site: PxxP_5
Protein A sequence: LSKPSPPLPPKRGIP
Protein A construct: Bio Ado Ado-LSKPSPPLPPKRGIP-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
The affinity was below the detection threshold of the assay.
Experiment series 7
Protein A protein: PHACTR4
Protein A fragment: 346-360
Protein A site: PxxP_6
Protein A sequence: PLPTHIPPEPPRTPP
Protein A construct: Bio Ado Ado-PLPTHIPPEPPRTPP-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 8
Protein A protein: PHACTR4
Protein A fragment: 353-367
Protein A site: PxxP_7
Protein A sequence: PEPPRTPPFPAKTFQ
Protein A construct: Bio Ado Ado-PEPPRTPPFPAKTFQ-CONH2
Protein B protein: PRMT2
Protein B fragment: 24-96
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: -0.01
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: 1
The affinity was below the detection threshold of the assay.
Experiment series 9
Protein A protein: PHACTR4
Protein A fragment: 400-414
Protein A site: PxxP_8
Protein A sequence: DQNFGEPHIPSRLPP
Protein A construct: Bio Ado Ado-DQNFGEPHIPSRLPP-CONH2
Protein B protein: PRMT2
Protein B fragment: 24-96
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3
Average holdup BI: -0.09
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.