Details of experiments between Q8WXE0 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: CASKIN2
Protein A fragment: 1-1202
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: 0.54
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): 4.42
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: 5.07

Experiment series 2

Protein A protein: CASKIN2
Protein A fragment: 1121-1135
Protein A site: PxxP_1
Protein A sequence: PRLGPRPVPPPRPES
Protein A construct: Bio Ado Ado-PRLGPRPVPPPRPES-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: CASKIN2
Protein A fragment: 683-697
Protein A site: PxxP_2
Protein A sequence: GGPEPLPLPPARSPS
Protein A construct: Bio Ado Ado-GGPEPLPLPPARSPS-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 4

Protein A protein: CASKIN2
Protein A fragment: 782-796
Protein A site: PxxP_3
Protein A sequence: AGPPATPPDPPRPKR
Protein A construct: Bio Ado Ado-AGPPATPPDPPRPKR-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
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: 3.64

Experiment series 5

Protein A protein: CASKIN2
Protein A fragment: 785-799
Protein A site: PxxP_4
Protein A sequence: PATPPDPPRPKRRSH
Protein A construct: Bio Ado Ado-PATPPDPPRPKRRSH-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 6

Protein A protein: CASKIN2
Protein A fragment: 862-876
Protein A site: PxxP_5
Protein A sequence: RARRKGPPPPPPKRL
Protein A construct: Bio Ado Ado-RARRKGPPPPPPKRL-CONH2

Protein B protein: PRMT2
Protein B fragment: 24-96
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3

Average holdup BI: 0.18
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: 4.11

Experiment series 7

Protein A protein: CASKIN2
Protein A fragment: 863-877
Protein A site: PxxP_6
Protein A sequence: ARRKGPPPPPPKRLS
Protein A construct: Bio Ado Ado-ARRKGPPPPPPKRLS-CONH2

Protein B protein: PRMT2
Protein B fragment: 24-96
Protein B site: SH3
Protein B construct: his6-MBP-TEVsite-SH3

Average holdup BI: 0.1
Holdup BI standard deviation: 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: 2
Measured pKd: 3.8

Experiment series 8

Protein A protein: CASKIN2
Protein A fragment: 864-878
Protein A site: PxxP_7
Protein A sequence: RRKGPPPPPPKRLSS
Protein A construct: Bio Ado Ado-RRKGPPPPPPKRLSS-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
Measured pKd: 3.73

Experiment series 9

Protein A protein: CASKIN2
Protein A fragment: 960-974
Protein A site: PxxP_8
Protein A sequence: IKQRPKPAGPPPRET
Protein A construct: Bio Ado Ado-IKQRPKPAGPPPRET-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 10

Protein A protein: CASKIN2
Protein A fragment: 969-983
Protein A site: PxxP_9
Protein A sequence: PPPRETPVPPGLDFN
Protein A construct: Bio Ado Ado-PPPRETPVPPGLDFN-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.