Interaction Details


Details of experiments between Q9BRD0 and O00499
Note that the results of all experiments are listed, regardless of the modification states of the fragments.


Experiment series 1

Protein A

Protein: BUD13
Fragment: 1-619
Construct: endogenous protein (Jurkat)

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: 0.66
Immobilized partner concentration (10-6M): 10
P value: 3.6
Experiment method: nHU_Jurkat
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-06-29
Number of measurements: 3
Measured pKd: 5.28
PUBMED ID: 38995680
Experimental Details

-log10(P Two-sided unpaired T-test (6 vs 3))

Experiment series 2

Protein A

Protein: BUD13
Fragment: 219-233
Site: PxxP_2
Sequence: RHDSPDPSPPRRARH
Construct: Biotinylated 15-mer peptide (crude)

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: 0.46
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022-06-29
Number of measurements: 2
Measured pKd: 4.72
PUBMED ID: 38995680

Experiment series 3

Protein A

Protein: BUD13
Fragment: 219-233
Site: PxxP_2
Sequence: RHDSPDPSPPRRARH
Construct: Biotinylated 15-mer peptide (crude)

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: 0.46
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022-06-29
Number of measurements: 2
Measured pKd: 4.72
PUBMED ID: N

Experiment series 4

Protein A

Protein: BUD13
Fragment: 219-233
Site: PxxP_2
Sequence: RHDSPDPSPPRRARH
Construct: Bio E E Ado Ado-RHDSPDPSPPRRARH-CONH2

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.44
Immobilized partner concentration (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023-01-16
Number of measurements: 1
Measured pKd: 4.68
PUBMED ID: 38995680

Experiment series 5

Protein A

Protein: BUD13
Fragment: 219-233
Site: PxxP_2
Sequence: RHDSPDPSPPRRARH
Construct: Biotinylated 15-mer peptide (crude)

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: 0.39
Holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022-10-24
Number of measurements: 2
Measured pKd: 4.59
PUBMED ID: 38995680

Experiment series 6

Protein A

Protein: BUD13
Fragment: 1-619
Construct: endogenous protein (Jurkat)

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: 0.2
Immobilized partner concentration (10-6M): 10
P value: 4.75
Experiment method: nHU_Jurkat
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-06-29
Number of measurements: 3
The affinity was below the detection threshold of the assay.
PUBMED ID: 38995680
Experimental Details

-log10(P Two-sided unpaired T-test (6 vs 3))

Experiment series 7

Protein A

Protein: BUD13
Fragment: 194-208
Site: PxxP_1
Sequence: RHDSPDPSPPRRPQH
Construct: Biotinylated 15-mer peptide (crude)

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: 0.02
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 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.
PUBMED ID: 38995680

Experiment series 8

Protein A

Protein: BUD13
Fragment: 194-208
Site: PxxP_1
Sequence: RHDSPDPSPPRRPQH
Construct: Biotinylated 15-mer peptide (crude)

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: 0.05
Holdup BI standard deviation: 0
Immobilized partner concentration (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.
PUBMED ID: 38995680

Experiment series 9

Protein A

Protein: BUD13
Fragment: 194-208
Site: PxxP_1
Sequence: RHDSPDPSPPRRPQH
Construct: Biotinylated 15-mer peptide (crude)

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: 0.05
Holdup BI standard deviation: 0
Immobilized partner concentration (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.
PUBMED ID: N

Experiment series 10

Protein A

Protein: BUD13
Fragment: 194-208
Site: PxxP_1
Sequence: RHDSPDPSPPRRPQH
Construct: Biotinylated 15-mer peptide (crude)

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: D537V
Construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: 0.01
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 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.
PUBMED ID: 38995680

Experiment series 11

Protein A

Protein: BUD13
Fragment: 219-233
Site: PxxP_2
Sequence: RHDSPDPSPPRRARH
Construct: Bio E E Ado Ado-RHDSPDPSPPRRARH-CONH2

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: D537V
Construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.03
Immobilized partner concentration (10-6M): 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.
PUBMED ID: 38995680

Experiment series 12

Protein A

Protein: BUD13
Fragment: 219-233
Site: PxxP_2
Sequence: RHDSPDPSPPRRARH
Construct: Biotinylated 15-mer peptide (crude)

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: D537V
Construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: 0.01
Holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 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.
PUBMED ID: 38995680

Experiment series 13

Protein A

Protein: BUD13
Fragment: 194-208
Site: PxxP_1
Sequence: RHDSPDPSPPRRPQH
Construct: Biotinylated 15-mer peptide (crude)

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: F584S
Construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: 0.02
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 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.
PUBMED ID: 38995680

Experiment series 14

Protein A

Protein: BUD13
Fragment: 219-233
Site: PxxP_2
Sequence: RHDSPDPSPPRRARH
Construct: Bio E E Ado Ado-RHDSPDPSPPRRARH-CONH2

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: F584S
Construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0
Immobilized partner concentration (10-6M): 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.
PUBMED ID: 38995680

Experiment series 15

Protein A

Protein: BUD13
Fragment: 219-233
Site: PxxP_2
Sequence: RHDSPDPSPPRRARH
Construct: Biotinylated 15-mer peptide (crude)

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: F584S
Construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: -0.1
Holdup BI standard deviation: 0.08
Immobilized partner concentration (10-6M): 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.
PUBMED ID: 38995680

Experiment series 16

Protein A

Protein: BUD13
Fragment: 219-233
Site: PxxP_2
Sequence: RHDSPDPSPPRRARH
Construct: Biotinylated 15-mer peptide (crude)

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: P551L
Construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: 0.16
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 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 pKd: 4.55
PUBMED ID: 38995680

Experiment series 17

Protein A

Protein: BUD13
Fragment: 219-233
Site: PxxP_2
Sequence: RHDSPDPSPPRRARH
Construct: Bio E E Ado Ado-RHDSPDPSPPRRARH-CONH2

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: P551L
Construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.17
Immobilized partner concentration (10-6M): 8
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023-01-17
Number of measurements: 1
Measured pKd: 4.43
PUBMED ID: 38995680

Experiment series 18

Protein A

Protein: BUD13
Fragment: 219-233
Site: PxxP_2
Sequence: RHDSPDPSPPRRARH
Construct: Biotinylated 15-mer peptide (crude)

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: P551L
Construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: 0.17
Holdup BI standard deviation: 0
Normalized holdup BI: 0.21
Normalized holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022-08-04
Number of measurements: 2
Measured pKd: 4.18
PUBMED ID: N

Experiment series 19

Protein A

Protein: BUD13
Fragment: 194-208
Site: PxxP_1
Sequence: RHDSPDPSPPRRPQH
Construct: Biotinylated 15-mer peptide (crude)

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: P551L
Construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: -0.04
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 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.
PUBMED ID: 38995680

Experiment series 20

Protein A

Protein: BUD13
Fragment: 194-208
Site: PxxP_1
Sequence: RHDSPDPSPPRRPQH
Construct: Biotinylated 15-mer peptide (crude)

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: P551L
Construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: -0.03
Holdup BI standard deviation: 0.06
Normalized holdup BI: -0.04
Normalized holdup BI standard deviation: 0.07
Immobilized partner concentration (10-6M): 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.
PUBMED ID: N

Experiment series 21

Protein A

Protein: BUD13
Fragment: 219-233
Site: PxxP_2
Sequence: RHDSPDPSPPRRARH
Construct: Biotinylated 15-mer peptide (crude)

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: Q540H
Construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: 0.46
Holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022-10-24
Number of measurements: 2
Measured pKd: 4.71
PUBMED ID: 38995680

Experiment series 22

Protein A

Protein: BUD13
Fragment: 219-233
Site: PxxP_2
Sequence: RHDSPDPSPPRRARH
Construct: Bio E E Ado Ado-RHDSPDPSPPRRARH-CONH2

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: Q540H
Construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.42
Immobilized partner concentration (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023-01-17
Number of measurements: 1
Measured pKd: 4.64
PUBMED ID: 38995680

Experiment series 23

Protein A

Protein: BUD13
Fragment: 219-233
Site: PxxP_2
Sequence: RHDSPDPSPPRRARH
Construct: Biotinylated 15-mer peptide (crude)

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: Q540H
Construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: 0.23
Holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022-08-04
Number of measurements: 2
Measured pKd: 4.23
PUBMED ID: N

Experiment series 24

Protein A

Protein: BUD13
Fragment: 194-208
Site: PxxP_1
Sequence: RHDSPDPSPPRRPQH
Construct: Biotinylated 15-mer peptide (crude)

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: Q540H
Construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: -0.02
Holdup BI standard deviation: 0.06
Immobilized partner concentration (10-6M): 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.
PUBMED ID: 38995680

Experiment series 25

Protein A

Protein: BUD13
Fragment: 194-208
Site: PxxP_1
Sequence: RHDSPDPSPPRRPQH
Construct: Biotinylated 15-mer peptide (crude)

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: Q540H
Construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: -0.01
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 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.
PUBMED ID: N

Experiment series 26

Protein A

Protein: BUD13
Fragment: 219-233
Site: PxxP_2
Sequence: RHDSPDPSPPRRARH
Construct: Biotinylated 15-mer peptide (crude)

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: R581C
Construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: 0.19
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 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 pKd: 4.64
PUBMED ID: 38995680

Experiment series 27

Protein A

Protein: BUD13
Fragment: 219-233
Site: PxxP_2
Sequence: RHDSPDPSPPRRARH
Construct: Bio E E Ado Ado-RHDSPDPSPPRRARH-CONH2

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: R581C
Construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.21
Immobilized partner concentration (10-6M): 10
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023-01-18
Number of measurements: 1
Measured pKd: 4.45
PUBMED ID: 38995680

Experiment series 28

Protein A

Protein: BUD13
Fragment: 219-233
Site: PxxP_2
Sequence: RHDSPDPSPPRRARH
Construct: Biotinylated 15-mer peptide (crude)

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: R581C
Construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: 0.2
Holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022-08-04
Number of measurements: 2
Measured pKd: 4.17
PUBMED ID: N

Experiment series 29

Protein A

Protein: BUD13
Fragment: 194-208
Site: PxxP_1
Sequence: RHDSPDPSPPRRPQH
Construct: Biotinylated 15-mer peptide (crude)

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: R581C
Construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: -0.03
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 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.
PUBMED ID: N

Experiment series 30

Protein A

Protein: BUD13
Fragment: 194-208
Site: PxxP_1
Sequence: RHDSPDPSPPRRPQH
Construct: Biotinylated 15-mer peptide (crude)

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: R581C
Construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: 0.01
Holdup BI standard deviation: 0.03
Immobilized partner concentration (10-6M): 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.
PUBMED ID: 38995680

Experiment series 31

Protein A

Protein: BUD13
Fragment: 219-233
Site: PxxP_2
Sequence: RHDSPDPSPPRRARH
Construct: Biotinylated 15-mer peptide (crude)

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: T532M
Construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: 0.43
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022-06-29
Number of measurements: 2
Measured pKd: 4.67
PUBMED ID: 38995680

Experiment series 32

Protein A

Protein: BUD13
Fragment: 219-233
Site: PxxP_2
Sequence: RHDSPDPSPPRRARH
Construct: Biotinylated 15-mer peptide (crude)

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: T532M
Construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: 0.43
Holdup BI standard deviation: 0.04
Immobilized partner concentration (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022-06-29
Number of measurements: 2
Measured pKd: 4.67
PUBMED ID: N

Experiment series 33

Protein A

Protein: BUD13
Fragment: 219-233
Site: PxxP_2
Sequence: RHDSPDPSPPRRARH
Construct: Biotinylated 15-mer peptide (crude)

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: T532M
Construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: 0.36
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022-10-24
Number of measurements: 2
Measured pKd: 4.54
PUBMED ID: 38995680

Experiment series 34

Protein A

Protein: BUD13
Fragment: 219-233
Site: PxxP_2
Sequence: RHDSPDPSPPRRARH
Construct: Bio E E Ado Ado-RHDSPDPSPPRRARH-CONH2

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: T532M
Construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.34
Immobilized partner concentration (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023-01-16
Number of measurements: 1
Measured pKd: 4.49
PUBMED ID: 38995680

Experiment series 35

Protein A

Protein: BUD13
Fragment: 194-208
Site: PxxP_1
Sequence: RHDSPDPSPPRRPQH
Construct: Biotinylated 15-mer peptide (crude)

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: T532M
Construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: 0.01
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 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.
PUBMED ID: 38995680

Experiment series 36

Protein A

Protein: BUD13
Fragment: 194-208
Site: PxxP_1
Sequence: RHDSPDPSPPRRPQH
Construct: Biotinylated 15-mer peptide (crude)

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: T532M
Construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: 0.02
Holdup BI standard deviation: 0.04
Immobilized partner concentration (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.
PUBMED ID: 38995680

Experiment series 37

Protein A

Protein: BUD13
Fragment: 194-208
Site: PxxP_1
Sequence: RHDSPDPSPPRRPQH
Construct: Biotinylated 15-mer peptide (crude)

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: T532M
Construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: 0.02
Holdup BI standard deviation: 0.04
Immobilized partner concentration (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.
PUBMED ID: N

Experiment series 38

Protein A

Protein: BUD13
Fragment: 219-233
Site: PxxP_2
Sequence: RHDSPDPSPPRRARH
Construct: Bio E E Ado Ado-RHDSPDPSPPRRARH-CONH2

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: V566M
Construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.37
Immobilized partner concentration (10-6M): 18
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023-01-18
Number of measurements: 1
Measured pKd: 4.54
PUBMED ID: 38995680

Experiment series 39

Protein A

Protein: BUD13
Fragment: 219-233
Site: PxxP_2
Sequence: RHDSPDPSPPRRARH
Construct: Biotinylated 15-mer peptide (crude)

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: V566M
Construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: 0.36
Holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022-10-24
Number of measurements: 2
Measured pKd: 4.53
PUBMED ID: 38995680

Experiment series 40

Protein A

Protein: BUD13
Fragment: 219-233
Site: PxxP_2
Sequence: RHDSPDPSPPRRARH
Construct: Biotinylated 15-mer peptide (crude)

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: V566M
Construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: 0.23
Holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022-08-04
Number of measurements: 2
Measured pKd: 4.23
PUBMED ID: N

Experiment series 41

Protein A

Protein: BUD13
Fragment: 194-208
Site: PxxP_1
Sequence: RHDSPDPSPPRRPQH
Construct: Biotinylated 15-mer peptide (crude)

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: V566M
Construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: 0
Holdup BI standard deviation: 0.02
Immobilized partner concentration (10-6M): 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.
PUBMED ID: 38995680

Experiment series 42

Protein A

Protein: BUD13
Fragment: 194-208
Site: PxxP_1
Sequence: RHDSPDPSPPRRPQH
Construct: Biotinylated 15-mer peptide (crude)

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: V566M
Construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: 0.01
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 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.
PUBMED ID: N

Experiment series 43

Protein A

Protein: BUD13
Fragment: 219-233
Site: PxxP_2
Sequence: RHDSPDPSPPRRARH
Construct: Biotinylated 15-mer peptide (crude)

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: V583I
Construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: 0.14
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 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 pKd: 4.28
PUBMED ID: 38995680

Experiment series 44

Protein A

Protein: BUD13
Fragment: 219-233
Site: PxxP_2
Sequence: RHDSPDPSPPRRARH
Construct: Bio E E Ado Ado-RHDSPDPSPPRRARH-CONH2

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: V583I
Construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.14
Immobilized partner concentration (10-6M): 13
Experiment method: HU (DAPF)
Details of affinity fitting: Quadratic binding equation
Date of measurement: 2023-01-17
Number of measurements: 1
Measured pKd: 4.13
PUBMED ID: 38995680

Experiment series 45

Protein A

Protein: BUD13
Fragment: 194-208
Site: PxxP_1
Sequence: RHDSPDPSPPRRPQH
Construct: Biotinylated 15-mer peptide (crude)

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: V583I
Construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: -0
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 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.
PUBMED ID: 38995680

Experiment series 46

Protein A

Protein: BUD13
Fragment: 194-208
Site: PxxP_1
Sequence: RHDSPDPSPPRRPQH
Construct: Biotinylated 15-mer peptide (crude)

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: Y531S
Construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: 0
Holdup BI standard deviation: 0.05
Immobilized partner concentration (10-6M): 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.
PUBMED ID: 38995680

Experiment series 47

Protein A

Protein: BUD13
Fragment: 219-233
Site: PxxP_2
Sequence: RHDSPDPSPPRRARH
Construct: Bio E E Ado Ado-RHDSPDPSPPRRARH-CONH2

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: Y531S
Construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.04
Immobilized partner concentration (10-6M): 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.
PUBMED ID: 38995680

Experiment series 48

Protein A

Protein: BUD13
Fragment: 219-233
Site: PxxP_2
Sequence: RHDSPDPSPPRRARH
Construct: Biotinylated 15-mer peptide (crude)

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: Y531S
Construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: 0.01
Holdup BI standard deviation: 0
Immobilized partner concentration (10-6M): 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.
PUBMED ID: 38995680