Details of experiments between  O00499  and P26358  Note that the results of all experiments are listed, regardless of the modification states of the fragments.
   Experiment series 1 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: DNMT1  Fragment: 1-1616  Construct: endogenous protein (Jurkat) 
Average holdup BI:  0.8 
Immobilized partner concentration (10-6 M):  10 
P value:  7.9 
Experiment method:  nHU_Jurkat 
Details of affinity fitting:  hyperbolic binding equation 
Date of measurement:  2022-06-29 
Number of measurements:  3 
Measured pK d : 5.6 
Experimental Details -log10(P Two-sided unpaired T-test (6 vs 3))
    Experiment series 2 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: DNMT1  Fragment: 1-1616  Construct: endogenous protein (Jurkat) 
Average holdup BI:  0.7 
Immobilized partner concentration (10-6 M):  10 
P value:  8.88 
Experiment method:  nHU_Jurkat 
Details of affinity fitting:  hyperbolic binding equation 
Date of measurement:  2022-06-29 
Number of measurements:  3 
Measured pK d : 5.36 
Experimental Details -log10(P Two-sided unpaired T-test (6 vs 3))
   Experiment series 3 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: DNMT1  Fragment: 129-143  Site: PxxP_1  Sequence: PKPLSKPRTPRRSKS  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.64 
Holdup BI standard deviation:  0.06 
Immobilized partner concentration (10-6 M):  18 
Experiment method:  HU (DAPF) 
Date of measurement:  2022-06-29 
Number of measurements:  2 
Measured pK d : 5.06 
   Experiment series 4 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: DNMT1  Fragment: 129-143  Site: PxxP_1  Sequence: PKPLSKPRTPRRSKS  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.64 
Holdup BI standard deviation:  0.06 
Immobilized partner concentration (10-6 M):  18 
Experiment method:  HU (DAPF) 
Date of measurement:  2022-06-29 
Number of measurements:  2 
Measured pK d : 5.06 
   Experiment series 5 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: DNMT1  Fragment: 129-143  Site: PxxP_1  Sequence: PKPLSKPRTPRRSKS  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.54 
Holdup BI standard deviation:  0.01 
Immobilized partner concentration (10-6 M):  18 
Experiment method:  HU (DAPF) 
Date of measurement:  2022-10-24 
Number of measurements:  2 
Measured pK d : 4.87 
   Experiment series 6 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Construct: his6-MBP-TEVsite-SH3 
Protein B Protein: DNMT1  Fragment: 129-143  Site: PxxP_1  Sequence: PKPLSKPRTPRRSKS  Construct: Bio E E Ado Ado-PKPLSKPRTPRRSKS-CONH2 
Average holdup BI:  0.11 
Immobilized partner concentration (10-6 M):  18 
Experiment method:  HU (DAPF) 
Details of affinity fitting:  Quadratic binding equation 
Date of measurement:  2023-01-16 
Number of measurements:  1 
Measured pK d : 3.84 
   Experiment series 7 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: DNMT1  Fragment: 323-337  Site: PxxP_2  Sequence: KRRKTTPKEPTEKKM  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.01 
Holdup BI standard deviation:  0.01 
Immobilized partner concentration (10-6 M):  18 
Experiment method:  HU (DAPF) 
Date of measurement:  2022-10-24 
Number of measurements:  2 
The affinity was below the detection threshold of the assay. 
   Experiment series 8 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: DNMT1  Fragment: 323-337  Site: PxxP_2  Sequence: KRRKTTPKEPTEKKM  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.04 
Holdup BI standard deviation:  0 
Immobilized partner concentration (10-6 M):  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: BIN1  Fragment: 513-593  Site: SH3  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: DNMT1  Fragment: 323-337  Site: PxxP_2  Sequence: KRRKTTPKEPTEKKM  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.04 
Holdup BI standard deviation:  0 
Immobilized partner concentration (10-6 M):  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: BIN1  Fragment: 513-593  Site: SH3  Modification: D537V  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: DNMT1  Fragment: 129-143  Site: PxxP_1  Sequence: PKPLSKPRTPRRSKS  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.08 
Holdup BI standard deviation:  0 
Immobilized partner concentration (10-6 M):  18 
Experiment method:  HU (DAPF) 
Date of measurement:  2022-10-24 
Number of measurements:  2 
Measured pK d : 3.67 
   Experiment series 11 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: D537V  Construct: his6-MBP-TEVsite-SH3 
Protein B Protein: DNMT1  Fragment: 129-143  Site: PxxP_1  Sequence: PKPLSKPRTPRRSKS  Construct: Bio E E Ado Ado-PKPLSKPRTPRRSKS-CONH2 
Average holdup BI:  -0.01 
Immobilized partner concentration (10-6 M):  18 
Experiment method:  HU (DAPF) 
Details of affinity fitting:  Quadratic binding equation 
Date of measurement:  2023-01-17 
Number of measurements:  1 
The affinity was below the detection threshold of the assay. 
   Experiment series 12 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: D537V  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: DNMT1  Fragment: 323-337  Site: PxxP_2  Sequence: KRRKTTPKEPTEKKM  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.01 
Holdup BI standard deviation:  0.02 
Immobilized partner concentration (10-6 M):  18 
Experiment method:  HU (DAPF) 
Date of measurement:  2022-10-24 
Number of measurements:  2 
The affinity was below the detection threshold of the assay. 
   Experiment series 13 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: F584S  Construct: his6-MBP-TEVsite-SH3 
Protein B Protein: DNMT1  Fragment: 129-143  Site: PxxP_1  Sequence: PKPLSKPRTPRRSKS  Construct: Bio E E Ado Ado-PKPLSKPRTPRRSKS-CONH2 
Average holdup BI:  -0.06 
Immobilized partner concentration (10-6 M):  18 
Experiment method:  HU (DAPF) 
Details of affinity fitting:  Quadratic binding equation 
Date of measurement:  2023-01-17 
Number of measurements:  1 
The affinity was below the detection threshold of the assay. 
   Experiment series 14 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: F584S  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: DNMT1  Fragment: 129-143  Site: PxxP_1  Sequence: PKPLSKPRTPRRSKS  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  -0 
Holdup BI standard deviation:  0.02 
Immobilized partner concentration (10-6 M):  18 
Experiment method:  HU (DAPF) 
Date of measurement:  2022-10-24 
Number of measurements:  2 
The affinity was below the detection threshold of the assay. 
   Experiment series 15 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: F584S  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: DNMT1  Fragment: 323-337  Site: PxxP_2  Sequence: KRRKTTPKEPTEKKM  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  -0 
Holdup BI standard deviation:  0.03 
Immobilized partner concentration (10-6 M):  18 
Experiment method:  HU (DAPF) 
Date of measurement:  2022-10-24 
Number of measurements:  2 
The affinity was below the detection threshold of the assay. 
   Experiment series 16 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: P551L  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: DNMT1  Fragment: 129-143  Site: PxxP_1  Sequence: PKPLSKPRTPRRSKS  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.27 
Holdup BI standard deviation:  0.02 
Immobilized partner concentration (10-6 M):  6 
Experiment method:  HU (DAPF) 
Details of affinity fitting:  Bait concentration was adjusted to fit the profile to WT 
Date of measurement:  2022-10-24 
Number of measurements:  2 
Measured pK d : 4.84 
   Experiment series 17 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: P551L  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: DNMT1  Fragment: 129-143  Site: PxxP_1  Sequence: PKPLSKPRTPRRSKS  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.29 
Holdup BI standard deviation:  0.02 
Normalized holdup BI:  0.36 
Normalized holdup BI standard deviation:  0.03 
Immobilized partner concentration (10-6 M):  18 
Experiment method:  HU (DAPF) 
Date of measurement:  2022-08-04 
Number of measurements:  2 
Measured pK d : 4.51 
   Experiment series 18 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: P551L  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: DNMT1  Fragment: 323-337  Site: PxxP_2  Sequence: KRRKTTPKEPTEKKM  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.05 
Holdup BI standard deviation:  0.03 
Immobilized partner concentration (10-6 M):  6 
Experiment method:  HU (DAPF) 
Details of affinity fitting:  Bait concentration was adjusted to fit the profile to WT 
Date of measurement:  2022-10-24 
Number of measurements:  2 
Measured pK d : 3.95 
   Experiment series 19 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: P551L  Construct: his6-MBP-TEVsite-SH3 
Protein B Protein: DNMT1  Fragment: 129-143  Site: PxxP_1  Sequence: PKPLSKPRTPRRSKS  Construct: Bio E E Ado Ado-PKPLSKPRTPRRSKS-CONH2 
Average holdup BI:  0.02 
Immobilized partner concentration (10-6 M):  8 
Experiment method:  HU (DAPF) 
Details of affinity fitting:  Quadratic binding equation 
Date of measurement:  2023-01-17 
Number of measurements:  1 
The affinity was below the detection threshold of the assay. 
   Experiment series 20 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: P551L  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: DNMT1  Fragment: 323-337  Site: PxxP_2  Sequence: KRRKTTPKEPTEKKM  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  -0 
Holdup BI standard deviation:  0.02 
Normalized holdup BI:  -0 
Normalized holdup BI standard deviation:  0.03 
Immobilized partner concentration (10-6 M):  18 
Experiment method:  HU (DAPF) 
Date of measurement:  2022-08-04 
Number of measurements:  2 
The affinity was below the detection threshold of the assay. 
   Experiment series 21 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: Q540H  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: DNMT1  Fragment: 129-143  Site: PxxP_1  Sequence: PKPLSKPRTPRRSKS  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.58 
Holdup BI standard deviation:  0.01 
Immobilized partner concentration (10-6 M):  18 
Experiment method:  HU (DAPF) 
Date of measurement:  2022-10-24 
Number of measurements:  2 
Measured pK d : 4.94 
   Experiment series 22 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: Q540H  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: DNMT1  Fragment: 129-143  Site: PxxP_1  Sequence: PKPLSKPRTPRRSKS  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.35 
Holdup BI standard deviation:  0.01 
Immobilized partner concentration (10-6 M):  18 
Experiment method:  HU (DAPF) 
Date of measurement:  2022-08-04 
Number of measurements:  2 
Measured pK d : 4.49 
   Experiment series 23 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: Q540H  Construct: his6-MBP-TEVsite-SH3 
Protein B Protein: DNMT1  Fragment: 129-143  Site: PxxP_1  Sequence: PKPLSKPRTPRRSKS  Construct: Bio E E Ado Ado-PKPLSKPRTPRRSKS-CONH2 
Average holdup BI:  0.14 
Immobilized partner concentration (10-6 M):  18 
Experiment method:  HU (DAPF) 
Details of affinity fitting:  Quadratic binding equation 
Date of measurement:  2023-01-17 
Number of measurements:  1 
Measured pK d : 3.99 
   Experiment series 24 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: Q540H  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: DNMT1  Fragment: 323-337  Site: PxxP_2  Sequence: KRRKTTPKEPTEKKM  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.05 
Holdup BI standard deviation:  0.02 
Immobilized partner concentration (10-6 M):  18 
Experiment method:  HU (DAPF) 
Date of measurement:  2022-10-24 
Number of measurements:  2 
Measured pK d : 3.49 
   Experiment series 25 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: Q540H  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: DNMT1  Fragment: 323-337  Site: PxxP_2  Sequence: KRRKTTPKEPTEKKM  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  -0 
Holdup BI standard deviation:  0.01 
Immobilized partner concentration (10-6 M):  18 
Experiment method:  HU (DAPF) 
Date of measurement:  2022-08-04 
Number of measurements:  2 
The affinity was below the detection threshold of the assay. 
   Experiment series 26 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: R581C  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: DNMT1  Fragment: 129-143  Site: PxxP_1  Sequence: PKPLSKPRTPRRSKS  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.3 
Holdup BI standard deviation:  0.01 
Immobilized partner concentration (10-6 M):  6 
Experiment method:  HU (DAPF) 
Details of affinity fitting:  Bait concentration was adjusted to fit the profile to WT 
Date of measurement:  2022-10-24 
Number of measurements:  2 
Measured pK d : 4.91 
   Experiment series 27 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: R581C  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: DNMT1  Fragment: 129-143  Site: PxxP_1  Sequence: PKPLSKPRTPRRSKS  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.34 
Holdup BI standard deviation:  0.01 
Immobilized partner concentration (10-6 M):  18 
Experiment method:  HU (DAPF) 
Date of measurement:  2022-08-04 
Number of measurements:  2 
Measured pK d : 4.5 
   Experiment series 28 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: R581C  Construct: his6-MBP-TEVsite-SH3 
Protein B Protein: DNMT1  Fragment: 129-143  Site: PxxP_1  Sequence: PKPLSKPRTPRRSKS  Construct: Bio E E Ado Ado-PKPLSKPRTPRRSKS-CONH2 
Average holdup BI:  -0.01 
Immobilized partner concentration (10-6 M):  10 
Experiment method:  HU (DAPF) 
Details of affinity fitting:  Quadratic binding equation 
Date of measurement:  2023-01-18 
Number of measurements:  1 
The affinity was below the detection threshold of the assay. 
   Experiment series 29 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: R581C  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: DNMT1  Fragment: 323-337  Site: PxxP_2  Sequence: KRRKTTPKEPTEKKM  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0 
Holdup BI standard deviation:  0.01 
Immobilized partner concentration (10-6 M):  18 
Experiment method:  HU (DAPF) 
Date of measurement:  2022-08-04 
Number of measurements:  2 
The affinity was below the detection threshold of the assay. 
   Experiment series 30 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: R581C  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: DNMT1  Fragment: 323-337  Site: PxxP_2  Sequence: KRRKTTPKEPTEKKM  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.02 
Holdup BI standard deviation:  0.02 
Immobilized partner concentration (10-6 M):  6 
Experiment method:  HU (DAPF) 
Details of affinity fitting:  Bait concentration was adjusted to fit the profile to WT 
Date of measurement:  2022-10-24 
Number of measurements:  2 
The affinity was below the detection threshold of the assay. 
   Experiment series 31 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: T532M  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: DNMT1  Fragment: 129-143  Site: PxxP_1  Sequence: PKPLSKPRTPRRSKS  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.57 
Holdup BI standard deviation:  0.04 
Immobilized partner concentration (10-6 M):  18 
Experiment method:  HU (DAPF) 
Date of measurement:  2022-06-29 
Number of measurements:  2 
Measured pK d : 4.92 
   Experiment series 32 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: T532M  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: DNMT1  Fragment: 129-143  Site: PxxP_1  Sequence: PKPLSKPRTPRRSKS  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.57 
Holdup BI standard deviation:  0.04 
Immobilized partner concentration (10-6 M):  18 
Experiment method:  HU (DAPF) 
Date of measurement:  2022-06-29 
Number of measurements:  2 
Measured pK d : 4.92 
   Experiment series 33 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: T532M  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: DNMT1  Fragment: 129-143  Site: PxxP_1  Sequence: PKPLSKPRTPRRSKS  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.5 
Holdup BI standard deviation:  0.02 
Immobilized partner concentration (10-6 M):  18 
Experiment method:  HU (DAPF) 
Date of measurement:  2022-10-24 
Number of measurements:  2 
Measured pK d : 4.8 
   Experiment series 34 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: T532M  Construct: his6-MBP-TEVsite-SH3 
Protein B Protein: DNMT1  Fragment: 129-143  Site: PxxP_1  Sequence: PKPLSKPRTPRRSKS  Construct: Bio E E Ado Ado-PKPLSKPRTPRRSKS-CONH2 
Average holdup BI:  0.13 
Immobilized partner concentration (10-6 M):  18 
Experiment method:  HU (DAPF) 
Details of affinity fitting:  Quadratic binding equation 
Date of measurement:  2023-01-16 
Number of measurements:  1 
Measured pK d : 3.93 
   Experiment series 35 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: T532M  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: DNMT1  Fragment: 323-337  Site: PxxP_2  Sequence: KRRKTTPKEPTEKKM  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.02 
Holdup BI standard deviation:  0.02 
Immobilized partner concentration (10-6 M):  18 
Experiment method:  HU (DAPF) 
Date of measurement:  2022-10-24 
Number of measurements:  2 
The affinity was below the detection threshold of the assay. 
   Experiment series 36 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: T532M  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: DNMT1  Fragment: 323-337  Site: PxxP_2  Sequence: KRRKTTPKEPTEKKM  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.05 
Holdup BI standard deviation:  0.01 
Immobilized partner concentration (10-6 M):  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 37 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: T532M  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: DNMT1  Fragment: 323-337  Site: PxxP_2  Sequence: KRRKTTPKEPTEKKM  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.05 
Holdup BI standard deviation:  0.01 
Immobilized partner concentration (10-6 M):  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 38 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: V566M  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: DNMT1  Fragment: 129-143  Site: PxxP_1  Sequence: PKPLSKPRTPRRSKS  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.52 
Holdup BI standard deviation:  0.02 
Immobilized partner concentration (10-6 M):  18 
Experiment method:  HU (DAPF) 
Date of measurement:  2022-10-24 
Number of measurements:  2 
Measured pK d : 4.83 
   Experiment series 39 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: V566M  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: DNMT1  Fragment: 129-143  Site: PxxP_1  Sequence: PKPLSKPRTPRRSKS  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.38 
Holdup BI standard deviation:  0.01 
Immobilized partner concentration (10-6 M):  18 
Experiment method:  HU (DAPF) 
Date of measurement:  2022-08-04 
Number of measurements:  2 
Measured pK d : 4.56 
   Experiment series 40 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: V566M  Construct: his6-MBP-TEVsite-SH3 
Protein B Protein: DNMT1  Fragment: 129-143  Site: PxxP_1  Sequence: PKPLSKPRTPRRSKS  Construct: Bio E E Ado Ado-PKPLSKPRTPRRSKS-CONH2 
Average holdup BI:  0.13 
Immobilized partner concentration (10-6 M):  18 
Experiment method:  HU (DAPF) 
Details of affinity fitting:  Quadratic binding equation 
Date of measurement:  2023-01-18 
Number of measurements:  1 
Measured pK d : 3.93 
   Experiment series 41 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: V566M  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: DNMT1  Fragment: 323-337  Site: PxxP_2  Sequence: KRRKTTPKEPTEKKM  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.02 
Holdup BI standard deviation:  0.01 
Immobilized partner concentration (10-6 M):  18 
Experiment method:  HU (DAPF) 
Date of measurement:  2022-08-04 
Number of measurements:  2 
The affinity was below the detection threshold of the assay. 
   Experiment series 42 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: V566M  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: DNMT1  Fragment: 323-337  Site: PxxP_2  Sequence: KRRKTTPKEPTEKKM  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.03 
Holdup BI standard deviation:  0.02 
Immobilized partner concentration (10-6 M):  18 
Experiment method:  HU (DAPF) 
Date of measurement:  2022-10-24 
Number of measurements:  2 
The affinity was below the detection threshold of the assay. 
   Experiment series 43 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: V583I  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: DNMT1  Fragment: 129-143  Site: PxxP_1  Sequence: PKPLSKPRTPRRSKS  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.42 
Holdup BI standard deviation:  0.02 
Immobilized partner concentration (10-6 M):  9 
Experiment method:  HU (DAPF) 
Details of affinity fitting:  Bait concentration was adjusted to fit the profile to WT 
Date of measurement:  2022-10-24 
Number of measurements:  2 
Measured pK d : 4.99 
   Experiment series 44 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: V583I  Construct: his6-MBP-TEVsite-SH3 
Protein B Protein: DNMT1  Fragment: 129-143  Site: PxxP_1  Sequence: PKPLSKPRTPRRSKS  Construct: Bio E E Ado Ado-PKPLSKPRTPRRSKS-CONH2 
Average holdup BI:  -0.01 
Immobilized partner concentration (10-6 M):  13 
Experiment method:  HU (DAPF) 
Details of affinity fitting:  Quadratic binding equation 
Date of measurement:  2023-01-17 
Number of measurements:  1 
The affinity was below the detection threshold of the assay. 
   Experiment series 45 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: V583I  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: DNMT1  Fragment: 323-337  Site: PxxP_2  Sequence: KRRKTTPKEPTEKKM  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.01 
Holdup BI standard deviation:  0 
Immobilized partner concentration (10-6 M):  9 
Experiment method:  HU (DAPF) 
Details of affinity fitting:  Bait concentration was adjusted to fit the profile to WT 
Date of measurement:  2022-10-24 
Number of measurements:  2 
The affinity was below the detection threshold of the assay. 
   Experiment series 46 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: Y531S  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: DNMT1  Fragment: 129-143  Site: PxxP_1  Sequence: PKPLSKPRTPRRSKS  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.23 
Holdup BI standard deviation:  0.01 
Immobilized partner concentration (10-6 M):  18 
Experiment method:  HU (DAPF) 
Date of measurement:  2022-10-24 
Number of measurements:  2 
Measured pK d : 4.24 
   Experiment series 47 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: Y531S  Construct: his6-MBP-TEVsite-SH3 
Protein B Protein: DNMT1  Fragment: 129-143  Site: PxxP_1  Sequence: PKPLSKPRTPRRSKS  Construct: Bio E E Ado Ado-PKPLSKPRTPRRSKS-CONH2 
Average holdup BI:  -0.01 
Immobilized partner concentration (10-6 M):  18 
Experiment method:  HU (DAPF) 
Details of affinity fitting:  Quadratic binding equation 
Date of measurement:  2023-01-18 
Number of measurements:  1 
The affinity was below the detection threshold of the assay. 
   Experiment series 48 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: Y531S  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: DNMT1  Fragment: 323-337  Site: PxxP_2  Sequence: KRRKTTPKEPTEKKM  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.02 
Holdup BI standard deviation:  0.02 
Immobilized partner concentration (10-6 M):  18 
Experiment method:  HU (DAPF) 
Date of measurement:  2022-10-24 
Number of measurements:  2 
The affinity was below the detection threshold of the assay. 
    
 
Please cite ProfAff! (Gogl, G. et al., Nature Communications 2022)  
© The ProfAff database is developped by the research team of Gergo Gogl .