Details of experiments between Q9UQ35 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: SRRM2
Protein A fragment: 1-2752
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: 0.15
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): 8.45
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: SRRM2
Protein A fragment: 1405-1419
Protein A site: PxxP_1
Protein A sequence: PVLDAVPRTPSRERS
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: -0.02
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 3
Protein A protein: SRRM2
Protein A fragment: 1426-1440
Protein A site: PxxP_2
Protein A sequence: EMKDGLPRTPSRRSR
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: 0.03
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 4
Protein A protein: SRRM2
Protein A fragment: 1464-1478
Protein A site: PxxP_3
Protein A sequence: PGMKDIPRTPSRGRS
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: 0.02
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 5
Protein A protein: SRRM2
Protein A fragment: 1484-1498
Protein A site: PxxP_4
Protein A sequence: PEPKALPQTPRPRSR
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: 0.06
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: 3.59
Experiment series 6
Protein A protein: SRRM2
Protein A fragment: 1653-1667
Protein A site: PxxP_5
Protein A sequence: SSTESSPEHPPKSRT
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: 2
The affinity was below the detection threshold of the assay.
Experiment series 7
Protein A protein: SRRM2
Protein A fragment: 2029-2043
Protein A site: PxxP_6
Protein A sequence: RSRSRTPLLPRKRSR
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: 2
Measured pKd: 4.11
Experiment series 8
Protein A protein: SRRM2
Protein A fragment: 2405-2419
Protein A site: PxxP_7
Protein A sequence: ARSRTPPSAPSQSRM
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: 0.02
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 9
Protein A protein: SRRM2
Protein A fragment: 2656-2670
Protein A site: PxxP_8
Protein A sequence: PQALPKPASPKKPPP
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
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 10
Protein A protein: SRRM2
Protein A fragment: 400-414
Protein A site: PxxP_9
Protein A sequence: TRDRSPPKSPEKLPQ
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: -0.02
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 11
Protein A protein: SRRM2
Protein A fragment: 975-989
Protein A site: PxxP_10
Protein A sequence: PDTKVKPETPPRQSH
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: 0.02
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.