Details of experiments between Q8IYB3 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: SRRM1
Protein A fragment: 1-904
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: 0.03
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): 1.17
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: SRRM1
Protein A fragment: 327-341
Protein A site: PxxP_1
Protein A sequence: TPPRRMPPPPRHRRS
Protein A construct: Biotinylated 15-mer peptide (crude)
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: 0.26
Holdup BI standard deviation: 0.06
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: 4.31
Experiment series 3
Protein A protein: SRRM1
Protein A fragment: 365-379
Protein A site: PxxP_2
Protein A sequence: SRSRSPPKKPPKRTS
Protein A construct: Biotinylated 15-mer peptide (crude)
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: 0.01
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: 4
The affinity was below the detection threshold of the assay.
Experiment series 4
Protein A protein: SRRM1
Protein A fragment: 395-409
Protein A site: PxxP_3
Protein A sequence: PRRRHRPSPPATPPP
Protein A construct: Biotinylated 15-mer peptide (crude)
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: 0.29
Holdup BI standard deviation: 0.05
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: 4.39
Experiment series 5
Protein A protein: SRRM1
Protein A fragment: 544-558
Protein A site: PxxP_4
Protein A sequence: GRRRRSPSPPPTRRR
Protein A construct: Biotinylated 15-mer peptide (crude)
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: 0.2
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: 4.16
Experiment series 6
Protein A protein: SRRM1
Protein A fragment: 557-571
Protein A site: PxxP_5
Protein A sequence: RRRSPSPAPPPRRRR
Protein A construct: Biotinylated 15-mer peptide (crude)
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: 0.18
Holdup BI standard deviation: 0.04
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: 4.11
Experiment series 7
Protein A protein: SRRM1
Protein A fragment: 567-581
Protein A site: PxxP_6
Protein A sequence: PRRRRTPTPPPRRRT
Protein A construct: Biotinylated 15-mer peptide (crude)
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: 0.18
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: 4
Measured pKd: 4.09
Experiment series 8
Protein A protein: SRRM1
Protein A fragment: 576-590
Protein A site: PxxP_7
Protein A sequence: PPRRRTPSPPPRRRS
Protein A construct: Biotinylated 15-mer peptide (crude)
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: 0.11
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: 4
Measured pKd: 3.84
Experiment series 9
Protein A protein: SRRM1
Protein A fragment: 600-614
Protein A site: PxxP_8
Protein A sequence: IQRRYSPSPPPKRRT
Protein A construct: Biotinylated 15-mer peptide (crude)
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: 0.05
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: 4
The affinity was below the detection threshold of the assay.
Experiment series 10
Protein A protein: SRRM1
Protein A fragment: 611-625
Protein A site: PxxP_9
Protein A sequence: KRRTASPPPPPKRRA
Protein A construct: Biotinylated 15-mer peptide (crude)
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: 0.12
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
Measured pKd: 3.91
Experiment series 11
Protein A protein: SRRM1
Protein A fragment: 612-626
Protein A site: PxxP_10
Protein A sequence: RRTASPPPPPKRRAS
Protein A construct: Biotinylated 15-mer peptide (crude)
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: 0.12
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
Measured pKd: 3.89
Experiment series 12
Protein A protein: SRRM1
Protein A fragment: 621-635
Protein A site: PxxP_11
Protein A sequence: PKRRASPSPPPKRRV
Protein A construct: Biotinylated 15-mer peptide (crude)
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: 0.04
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: 4
The affinity was below the detection threshold of the assay.
Experiment series 13
Protein A protein: SRRM1
Protein A fragment: 759-773
Protein A site: PxxP_12
Protein A sequence: PAAKKPPAPPSPVQS
Protein A construct: Biotinylated 15-mer peptide (crude)
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: 0.03
Holdup BI standard deviation: 0
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 14
Protein A protein: SRRM1
Protein A fragment: 776-790
Protein A site: PxxP_13
Protein A sequence: PSTNWSPAVPVKKAK
Protein A construct: Biotinylated 15-mer peptide (crude)
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
Holdup BI standard deviation: 0
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 15
Protein A protein: SRRM1
Protein A fragment: 576-590
Protein A site: PxxP_7
Protein A sequence: PPRRRTPSPPPRRRS
Protein A construct: Bio E E Ado Ado-PPRRRTPSPPPRRRS-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.