Details of experiments between  O00499  and P06400  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: RB1  Fragment: 1-928  Construct: endogenous protein (Jurkat) 
Average holdup BI:  0.44 
Immobilized partner concentration (10-6 M):  10 
P value:  6.01 
Experiment method:  nHU_Jurkat 
Details of affinity fitting:  hyperbolic binding equation 
Date of measurement:  2022-06-29 
Number of measurements:  3 
Measured pK d : 4.9 
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: RB1  Fragment: 1-928  Construct: endogenous protein (Jurkat) 
Average holdup BI:  0.3 
Immobilized partner concentration (10-6 M):  10 
P value:  7.92 
Experiment method:  nHU_Jurkat 
Details of affinity fitting:  hyperbolic binding equation 
Date of measurement:  2022-06-29 
Number of measurements:  3 
Measured pK d : 4.63 
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: RB1  Fragment: 862-876  Site: PxxP_5  Sequence: SAEGSNPPKPLKKLR  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.08 
Holdup BI standard deviation:  0.02 
Immobilized partner concentration (10-6 M):  18 
Experiment method:  HU (DAPF) 
Date of measurement:  2022-06-29 
Number of measurements:  2 
Measured pK d : 3.67 
   Experiment series 4 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 862-876  Site: PxxP_5  Sequence: SAEGSNPPKPLKKLR  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.08 
Holdup BI standard deviation:  0.02 
Immobilized partner concentration (10-6 M):  18 
Experiment method:  HU (DAPF) 
Date of measurement:  2022-06-29 
Number of measurements:  2 
Measured pK d : 3.67 
   Experiment series 5 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 244-258  Site: PxxP_1  Sequence: IPINGSPRTPRRGQN  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.07 
Holdup BI standard deviation:  0.03 
Immobilized partner concentration (10-6 M):  18 
Experiment method:  HU (DAPF) 
Date of measurement:  2022-06-29 
Number of measurements:  2 
Measured pK d : 3.61 
   Experiment series 6 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 244-258  Site: PxxP_1  Sequence: IPINGSPRTPRRGQN  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.07 
Holdup BI standard deviation:  0.03 
Immobilized partner concentration (10-6 M):  18 
Experiment method:  HU (DAPF) 
Date of measurement:  2022-06-29 
Number of measurements:  2 
Measured pK d : 3.61 
   Experiment series 7 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 244-258  Site: PxxP_1  Sequence: IPINGSPRTPRRGQN  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.06 
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 : 3.57 
   Experiment series 8 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 780-794  Site: PxxP_3  Sequence: SPIPHIPRSPYKFPS  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.06 
Holdup BI standard deviation:  0.03 
Immobilized partner concentration (10-6 M):  18 
Experiment method:  HU (DAPF) 
Date of measurement:  2022-06-29 
Number of measurements:  2 
Measured pK d : 3.55 
   Experiment series 9 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 780-794  Site: PxxP_3  Sequence: SPIPHIPRSPYKFPS  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.06 
Holdup BI standard deviation:  0.03 
Immobilized partner concentration (10-6 M):  18 
Experiment method:  HU (DAPF) 
Date of measurement:  2022-06-29 
Number of measurements:  2 
Measured pK d : 3.55 
   Experiment series 10 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 365-379  Site: PxxP_2  Sequence: EVNVIPPHTPVRTVM  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  -0.01 
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 
The affinity was below the detection threshold of the assay. 
   Experiment series 11 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 365-379  Site: PxxP_2  Sequence: EVNVIPPHTPVRTVM  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  -0.01 
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 12 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 365-379  Site: PxxP_2  Sequence: EVNVIPPHTPVRTVM  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  -0.01 
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 13 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 780-794  Site: PxxP_3  Sequence: SPIPHIPRSPYKFPS  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.04 
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 14 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 787-801  Site: PxxP_4  Sequence: RSPYKFPSSPLRIPG  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 15 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 787-801  Site: PxxP_4  Sequence: RSPYKFPSSPLRIPG  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:  2022-06-29 
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  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 787-801  Site: PxxP_4  Sequence: RSPYKFPSSPLRIPG  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:  2022-06-29 
Number of measurements:  2 
The affinity was below the detection threshold of the assay. 
   Experiment series 17 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 862-876  Site: PxxP_5  Sequence: SAEGSNPPKPLKKLR  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.04 
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 18 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: D537V  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 244-258  Site: PxxP_1  Sequence: IPINGSPRTPRRGQN  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  -0.01 
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 19 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: D537V  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 365-379  Site: PxxP_2  Sequence: EVNVIPPHTPVRTVM  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  -0.04 
Holdup BI standard deviation:  0.04 
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 20 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: D537V  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 780-794  Site: PxxP_3  Sequence: SPIPHIPRSPYKFPS  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 21 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: D537V  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 787-801  Site: PxxP_4  Sequence: RSPYKFPSSPLRIPG  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:  2022-10-24 
Number of measurements:  2 
The affinity was below the detection threshold of the assay. 
   Experiment series 22 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: D537V  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 862-876  Site: PxxP_5  Sequence: SAEGSNPPKPLKKLR  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 23 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: F584S  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 244-258  Site: PxxP_1  Sequence: IPINGSPRTPRRGQN  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 24 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: F584S  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 365-379  Site: PxxP_2  Sequence: EVNVIPPHTPVRTVM  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 25 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: F584S  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 780-794  Site: PxxP_3  Sequence: SPIPHIPRSPYKFPS  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  -0.04 
Holdup BI standard deviation:  0.06 
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 26 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: F584S  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 787-801  Site: PxxP_4  Sequence: RSPYKFPSSPLRIPG  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 27 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: F584S  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 862-876  Site: PxxP_5  Sequence: SAEGSNPPKPLKKLR  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 28 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: P551L  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 244-258  Site: PxxP_1  Sequence: IPINGSPRTPRRGQN  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.06 
Holdup BI standard deviation:  0.07 
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 : 4.06 
   Experiment series 29 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: P551L  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 862-876  Site: PxxP_5  Sequence: SAEGSNPPKPLKKLR  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.06 
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 : 4.04 
   Experiment series 30 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: P551L  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 244-258  Site: PxxP_1  Sequence: IPINGSPRTPRRGQN  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.07 
Holdup BI standard deviation:  0.11 
Normalized holdup BI:  0.08 
Normalized holdup BI standard deviation:  0.14 
Immobilized partner concentration (10-6 M):  18 
Experiment method:  HU (DAPF) 
Date of measurement:  2022-08-04 
Number of measurements:  2 
Measured pK d : 3.71 
   Experiment series 31 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: P551L  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 365-379  Site: PxxP_2  Sequence: EVNVIPPHTPVRTVM  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.02 
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 
The affinity was below the detection threshold of the assay. 
   Experiment series 32 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: P551L  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 365-379  Site: PxxP_2  Sequence: EVNVIPPHTPVRTVM  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.02 
Holdup BI standard deviation:  0.01 
Normalized holdup BI:  0.02 
Normalized holdup BI standard deviation:  0.02 
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 33 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: P551L  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 780-794  Site: PxxP_3  Sequence: SPIPHIPRSPYKFPS  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.02 
Holdup BI standard deviation:  0 
Normalized holdup BI:  0.03 
Normalized 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 34 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: P551L  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 780-794  Site: PxxP_3  Sequence: SPIPHIPRSPYKFPS  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.03 
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 
The affinity was below the detection threshold of the assay. 
   Experiment series 35 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: P551L  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 787-801  Site: PxxP_4  Sequence: RSPYKFPSSPLRIPG  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 36 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: P551L  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 787-801  Site: PxxP_4  Sequence: RSPYKFPSSPLRIPG  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.03 
Holdup BI standard deviation:  0 
Normalized holdup BI:  0.04 
Normalized holdup BI standard deviation:  0 
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 37 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: P551L  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 862-876  Site: PxxP_5  Sequence: SAEGSNPPKPLKKLR  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.05 
Holdup BI standard deviation:  0.01 
Normalized holdup BI:  0.06 
Normalized 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 38 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: Q540H  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 244-258  Site: PxxP_1  Sequence: IPINGSPRTPRRGQN  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.11 
Holdup BI standard deviation:  0.04 
Immobilized partner concentration (10-6 M):  18 
Experiment method:  HU (DAPF) 
Date of measurement:  2022-10-24 
Number of measurements:  2 
Measured pK d : 3.84 
   Experiment series 39 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: Q540H  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 244-258  Site: PxxP_1  Sequence: IPINGSPRTPRRGQN  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.1 
Holdup BI standard deviation:  0.1 
Immobilized partner concentration (10-6 M):  18 
Experiment method:  HU (DAPF) 
Date of measurement:  2022-08-04 
Number of measurements:  2 
Measured pK d : 3.78 
   Experiment series 40 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: Q540H  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 862-876  Site: PxxP_5  Sequence: SAEGSNPPKPLKKLR  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.05 
Holdup BI standard deviation:  0.05 
Immobilized partner concentration (10-6 M):  18 
Experiment method:  HU (DAPF) 
Date of measurement:  2022-10-24 
Number of measurements:  2 
Measured pK d : 3.47 
   Experiment series 41 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: Q540H  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 365-379  Site: PxxP_2  Sequence: EVNVIPPHTPVRTVM  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.02 
Holdup BI standard deviation:  0 
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: Q540H  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 365-379  Site: PxxP_2  Sequence: EVNVIPPHTPVRTVM  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: Q540H  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 780-794  Site: PxxP_3  Sequence: SPIPHIPRSPYKFPS  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  -0.01 
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 44 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: Q540H  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 780-794  Site: PxxP_3  Sequence: SPIPHIPRSPYKFPS  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.03 
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 45 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: Q540H  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 787-801  Site: PxxP_4  Sequence: RSPYKFPSSPLRIPG  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  -0.01 
Holdup BI standard deviation:  0.04 
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 46 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: Q540H  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 787-801  Site: PxxP_4  Sequence: RSPYKFPSSPLRIPG  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.02 
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 47 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: Q540H  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 862-876  Site: PxxP_5  Sequence: SAEGSNPPKPLKKLR  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:  2022-08-04 
Number of measurements:  2 
The affinity was below the detection threshold of the assay. 
   Experiment series 48 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: R581C  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 244-258  Site: PxxP_1  Sequence: IPINGSPRTPRRGQN  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.06 
Holdup BI standard deviation:  0.05 
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 : 4.03 
   Experiment series 49 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: R581C  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 244-258  Site: PxxP_1  Sequence: IPINGSPRTPRRGQN  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.07 
Holdup BI standard deviation:  0.09 
Immobilized partner concentration (10-6 M):  18 
Experiment method:  HU (DAPF) 
Date of measurement:  2022-08-04 
Number of measurements:  2 
Measured pK d : 3.65 
   Experiment series 50 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: R581C  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 365-379  Site: PxxP_2  Sequence: EVNVIPPHTPVRTVM  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  -0.01 
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 
The affinity was below the detection threshold of the assay. 
   Experiment series 51 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: R581C  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 365-379  Site: PxxP_2  Sequence: EVNVIPPHTPVRTVM  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:  2022-08-04 
Number of measurements:  2 
The affinity was below the detection threshold of the assay. 
   Experiment series 52 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: R581C  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 780-794  Site: PxxP_3  Sequence: SPIPHIPRSPYKFPS  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 53 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: R581C  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 780-794  Site: PxxP_3  Sequence: SPIPHIPRSPYKFPS  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 54 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: R581C  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 787-801  Site: PxxP_4  Sequence: RSPYKFPSSPLRIPG  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:  2022-08-04 
Number of measurements:  2 
The affinity was below the detection threshold of the assay. 
   Experiment series 55 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: R581C  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 787-801  Site: PxxP_4  Sequence: RSPYKFPSSPLRIPG  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0 
Holdup BI standard deviation:  0 
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 56 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: R581C  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 862-876  Site: PxxP_5  Sequence: SAEGSNPPKPLKKLR  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.01 
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 57 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: R581C  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 862-876  Site: PxxP_5  Sequence: SAEGSNPPKPLKKLR  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.04 
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 
The affinity was below the detection threshold of the assay. 
   Experiment series 58 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: T532M  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 862-876  Site: PxxP_5  Sequence: SAEGSNPPKPLKKLR  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.07 
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 
Measured pK d : 3.63 
   Experiment series 59 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: T532M  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 244-258  Site: PxxP_1  Sequence: IPINGSPRTPRRGQN  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.07 
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 
Measured pK d : 3.62 
   Experiment series 60 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: T532M  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 244-258  Site: PxxP_1  Sequence: IPINGSPRTPRRGQN  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.04 
Holdup BI standard deviation:  0.05 
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 61 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: T532M  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 244-258  Site: PxxP_1  Sequence: IPINGSPRTPRRGQN  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.04 
Holdup BI standard deviation:  0.05 
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 62 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: T532M  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 365-379  Site: PxxP_2  Sequence: EVNVIPPHTPVRTVM  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  -0.02 
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 
The affinity was below the detection threshold of the assay. 
   Experiment series 63 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: T532M  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 365-379  Site: PxxP_2  Sequence: EVNVIPPHTPVRTVM  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.03 
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 64 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: T532M  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 365-379  Site: PxxP_2  Sequence: EVNVIPPHTPVRTVM  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.03 
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 65 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: T532M  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 780-794  Site: PxxP_3  Sequence: SPIPHIPRSPYKFPS  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 66 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: T532M  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 780-794  Site: PxxP_3  Sequence: SPIPHIPRSPYKFPS  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:  2022-06-29 
Number of measurements:  2 
The affinity was below the detection threshold of the assay. 
   Experiment series 67 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: T532M  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 780-794  Site: PxxP_3  Sequence: SPIPHIPRSPYKFPS  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:  2022-06-29 
Number of measurements:  2 
The affinity was below the detection threshold of the assay. 
   Experiment series 68 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: T532M  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 787-801  Site: PxxP_4  Sequence: RSPYKFPSSPLRIPG  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  -0.02 
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 
The affinity was below the detection threshold of the assay. 
   Experiment series 69 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: T532M  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 787-801  Site: PxxP_4  Sequence: RSPYKFPSSPLRIPG  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  -0.02 
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 
The affinity was below the detection threshold of the assay. 
   Experiment series 70 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: T532M  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 787-801  Site: PxxP_4  Sequence: RSPYKFPSSPLRIPG  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 71 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: T532M  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 862-876  Site: PxxP_5  Sequence: SAEGSNPPKPLKKLR  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.02 
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 
The affinity was below the detection threshold of the assay. 
   Experiment series 72 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: T532M  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 862-876  Site: PxxP_5  Sequence: SAEGSNPPKPLKKLR  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.02 
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 
The affinity was below the detection threshold of the assay. 
   Experiment series 73 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: V566M  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 244-258  Site: PxxP_1  Sequence: IPINGSPRTPRRGQN  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.09 
Holdup BI standard deviation:  0.07 
Immobilized partner concentration (10-6 M):  18 
Experiment method:  HU (DAPF) 
Date of measurement:  2022-08-04 
Number of measurements:  2 
Measured pK d : 3.74 
   Experiment series 74 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: V566M  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 862-876  Site: PxxP_5  Sequence: SAEGSNPPKPLKKLR  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.07 
Holdup BI standard deviation:  0.05 
Immobilized partner concentration (10-6 M):  18 
Experiment method:  HU (DAPF) 
Date of measurement:  2022-10-24 
Number of measurements:  2 
Measured pK d : 3.62 
   Experiment series 75 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: V566M  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 862-876  Site: PxxP_5  Sequence: SAEGSNPPKPLKKLR  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.07 
Holdup BI standard deviation:  0.05 
Immobilized partner concentration (10-6 M):  18 
Experiment method:  HU (DAPF) 
Date of measurement:  2022-08-04 
Number of measurements:  2 
Measured pK d : 3.62 
   Experiment series 76 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: V566M  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 244-258  Site: PxxP_1  Sequence: IPINGSPRTPRRGQN  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.06 
Holdup BI standard deviation:  0.06 
Immobilized partner concentration (10-6 M):  18 
Experiment method:  HU (DAPF) 
Date of measurement:  2022-10-24 
Number of measurements:  2 
Measured pK d : 3.57 
   Experiment series 77 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: V566M  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 365-379  Site: PxxP_2  Sequence: EVNVIPPHTPVRTVM  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:  2022-08-04 
Number of measurements:  2 
The affinity was below the detection threshold of the assay. 
   Experiment series 78 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: V566M  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 365-379  Site: PxxP_2  Sequence: EVNVIPPHTPVRTVM  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 79 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: V566M  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 780-794  Site: PxxP_3  Sequence: SPIPHIPRSPYKFPS  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.01 
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 80 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: V566M  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 780-794  Site: PxxP_3  Sequence: SPIPHIPRSPYKFPS  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.02 
Holdup BI standard deviation:  0 
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 81 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: V566M  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 787-801  Site: PxxP_4  Sequence: RSPYKFPSSPLRIPG  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:  2022-08-04 
Number of measurements:  2 
The affinity was below the detection threshold of the assay. 
   Experiment series 82 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: V566M  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 787-801  Site: PxxP_4  Sequence: RSPYKFPSSPLRIPG  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 83 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: V583I  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 244-258  Site: PxxP_1  Sequence: IPINGSPRTPRRGQN  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.02 
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 
The affinity was below the detection threshold of the assay. 
   Experiment series 84 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: V583I  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 365-379  Site: PxxP_2  Sequence: EVNVIPPHTPVRTVM  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  -0.01 
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 
The affinity was below the detection threshold of the assay. 
   Experiment series 85 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: V583I  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 780-794  Site: PxxP_3  Sequence: SPIPHIPRSPYKFPS  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  -0.06 
Holdup BI standard deviation:  0.03 
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 86 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: V583I  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 787-801  Site: PxxP_4  Sequence: RSPYKFPSSPLRIPG  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.03 
Holdup BI standard deviation:  0.03 
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 87 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: V583I  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 862-876  Site: PxxP_5  Sequence: SAEGSNPPKPLKKLR  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.02 
Holdup BI standard deviation:  0.05 
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 88 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: Y531S  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 244-258  Site: PxxP_1  Sequence: IPINGSPRTPRRGQN  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 89 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: Y531S  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 365-379  Site: PxxP_2  Sequence: EVNVIPPHTPVRTVM  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  -0 
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 
The affinity was below the detection threshold of the assay. 
   Experiment series 90 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: Y531S  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 780-794  Site: PxxP_3  Sequence: SPIPHIPRSPYKFPS  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.02 
Holdup BI standard deviation:  0.04 
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 91 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: Y531S  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 787-801  Site: PxxP_4  Sequence: RSPYKFPSSPLRIPG  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 92 Protein A Protein: BIN1  Fragment: 513-593  Site: SH3  Modification: Y531S  Construct: avi(biotin)-his6-MBP-TEVsite-SH3 
Protein B Protein: RB1  Fragment: 862-876  Site: PxxP_5  Sequence: SAEGSNPPKPLKKLR  Construct: Biotinylated 15-mer peptide (crude) 
Average holdup BI:  0.03 
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. 
    
 
Please cite ProfAff! (Gogl, G. et al., Nature Communications 2022)  
© The ProfAff database is developped by the research team of Gergo Gogl .