Interaction Details


Details of experiments between A6NHR9 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: SMCHD1
Fragment: 1-2005
Construct: endogenous protein (Jurkat)

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: 0.87
Immobilized partner concentration (10-6M): 10
P value: 6.57
Experiment method: nHU_Jurkat
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-06-29
Number of measurements: 3
Measured pKd: 5.84
PUBMED ID: 38995680
Experimental Details

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

Experiment series 2

Protein A

Protein: SMCHD1
Fragment: 1977-1991
Site: PxxP_2
Sequence: VETTDCPVPPKRMRR
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: 0.88
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022-06-29
Number of measurements: 2
Measured pKd: 5.69
PUBMED ID: 38995680

Experiment series 3

Protein A

Protein: SMCHD1
Fragment: 1977-1991
Site: PxxP_2
Sequence: VETTDCPVPPKRMRR
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: 0.88
Holdup BI standard deviation: 0.01
Immobilized partner concentration (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022-06-29
Number of measurements: 2
Measured pKd: 5.69
PUBMED ID: N

Experiment series 4

Protein A

Protein: SMCHD1
Fragment: 1977-1991
Site: PxxP_2
Sequence: VETTDCPVPPKRMRR
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: 0.83
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: 5.51
PUBMED ID: 38995680

Experiment series 5

Protein A

Protein: SMCHD1
Fragment: 1-2005
Construct: endogenous protein (Jurkat)

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: 0.69
Immobilized partner concentration (10-6M): 10
P value: 8.16
Experiment method: nHU_Jurkat
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-06-29
Number of measurements: 3
Measured pKd: 5.35
PUBMED ID: 38995680
Experimental Details

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

Experiment series 6

Protein A

Protein: SMCHD1
Fragment: 1977-1991
Site: PxxP_2
Sequence: VETTDCPVPPKRMRR
Construct: Bio E E Ado Ado-VETTDCPVPPKRMRR-CONH2

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.47
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: 4.74
PUBMED ID: 38995680

Experiment series 7

Protein A

Protein: SMCHD1
Fragment: 1139-1153
Site: PxxP_1
Sequence: FTVRPLPDEPKHLKC
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: -0.01
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 8

Protein A

Protein: SMCHD1
Fragment: 1139-1153
Site: PxxP_1
Sequence: FTVRPLPDEPKHLKC
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: 0.03
Holdup BI standard deviation: 0.02
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: SMCHD1
Fragment: 1139-1153
Site: PxxP_1
Sequence: FTVRPLPDEPKHLKC
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: 0.03
Holdup BI standard deviation: 0.02
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: SMCHD1
Fragment: 437-451
Site: PxxP_3
Sequence: ETYPDDPCFPSKLKD
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
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: SMCHD1
Fragment: 437-451
Site: PxxP_3
Sequence: ETYPDDPCFPSKLKD
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: 0.01
Holdup BI standard deviation: 0.02
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 12

Protein A

Protein: SMCHD1
Fragment: 437-451
Site: PxxP_3
Sequence: ETYPDDPCFPSKLKD
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: 0.01
Holdup BI standard deviation: 0.02
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 13

Protein A

Protein: SMCHD1
Fragment: 1977-1991
Site: PxxP_2
Sequence: VETTDCPVPPKRMRR
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: 0.43
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
Measured pKd: 4.67
PUBMED ID: 38995680

Experiment series 14

Protein A

Protein: SMCHD1
Fragment: 1977-1991
Site: PxxP_2
Sequence: VETTDCPVPPKRMRR
Construct: Bio E E Ado Ado-VETTDCPVPPKRMRR-CONH2

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: D537V
Construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.11
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: 3.87
PUBMED ID: 38995680

Experiment series 15

Protein A

Protein: SMCHD1
Fragment: 1139-1153
Site: PxxP_1
Sequence: FTVRPLPDEPKHLKC
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
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 16

Protein A

Protein: SMCHD1
Fragment: 437-451
Site: PxxP_3
Sequence: ETYPDDPCFPSKLKD
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
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 17

Protein A

Protein: SMCHD1
Fragment: 1139-1153
Site: PxxP_1
Sequence: FTVRPLPDEPKHLKC
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: -0.07
Holdup BI standard deviation: 0.1
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 18

Protein A

Protein: SMCHD1
Fragment: 1977-1991
Site: PxxP_2
Sequence: VETTDCPVPPKRMRR
Construct: Bio E E Ado Ado-VETTDCPVPPKRMRR-CONH2

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: F584S
Construct: his6-MBP-TEVsite-SH3
Average holdup BI: -0.15
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 19

Protein A

Protein: SMCHD1
Fragment: 1977-1991
Site: PxxP_2
Sequence: VETTDCPVPPKRMRR
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: -0.01
Holdup BI standard deviation: 0.07
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 20

Protein A

Protein: SMCHD1
Fragment: 437-451
Site: PxxP_3
Sequence: ETYPDDPCFPSKLKD
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: -0.05
Holdup BI standard deviation: 0.11
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 21

Protein A

Protein: SMCHD1
Fragment: 1977-1991
Site: PxxP_2
Sequence: VETTDCPVPPKRMRR
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: 0.64
Holdup BI standard deviation: 0
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: 5.62
PUBMED ID: 38995680

Experiment series 22

Protein A

Protein: SMCHD1
Fragment: 1977-1991
Site: PxxP_2
Sequence: VETTDCPVPPKRMRR
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: 0.57
Holdup BI standard deviation: 0.01
Normalized holdup BI: 0.72
Normalized 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
Measured pKd: 5.18
PUBMED ID: N

Experiment series 23

Protein A

Protein: SMCHD1
Fragment: 1977-1991
Site: PxxP_2
Sequence: VETTDCPVPPKRMRR
Construct: Bio E E Ado Ado-VETTDCPVPPKRMRR-CONH2

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: P551L
Construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.31
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: 4.79
PUBMED ID: 38995680

Experiment series 24

Protein A

Protein: SMCHD1
Fragment: 1139-1153
Site: PxxP_1
Sequence: FTVRPLPDEPKHLKC
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: 0.01
Holdup BI standard deviation: 0.03
Normalized holdup BI: 0.01
Normalized holdup BI standard deviation: 0.04
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 25

Protein A

Protein: SMCHD1
Fragment: 1139-1153
Site: PxxP_1
Sequence: FTVRPLPDEPKHLKC
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: 0.01
Holdup BI standard deviation: 0.02
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 26

Protein A

Protein: SMCHD1
Fragment: 437-451
Site: PxxP_3
Sequence: ETYPDDPCFPSKLKD
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: -0.01
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 27

Protein A

Protein: SMCHD1
Fragment: 437-451
Site: PxxP_3
Sequence: ETYPDDPCFPSKLKD
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: 0.02
Holdup BI standard deviation: 0
Normalized holdup BI: 0.02
Normalized 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
The affinity was below the detection threshold of the assay.
PUBMED ID: N

Experiment series 28

Protein A

Protein: SMCHD1
Fragment: 1977-1991
Site: PxxP_2
Sequence: VETTDCPVPPKRMRR
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: 0.79
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: 5.4
PUBMED ID: 38995680

Experiment series 29

Protein A

Protein: SMCHD1
Fragment: 1977-1991
Site: PxxP_2
Sequence: VETTDCPVPPKRMRR
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: 0.64
Holdup BI standard deviation: 0.1
Immobilized partner concentration (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022-08-04
Number of measurements: 2
Measured pKd: 5.04
PUBMED ID: N

Experiment series 30

Protein A

Protein: SMCHD1
Fragment: 1977-1991
Site: PxxP_2
Sequence: VETTDCPVPPKRMRR
Construct: Bio E E Ado Ado-VETTDCPVPPKRMRR-CONH2

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: Q540H
Construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.52
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: 4.82
PUBMED ID: 38995680

Experiment series 31

Protein A

Protein: SMCHD1
Fragment: 437-451
Site: PxxP_3
Sequence: ETYPDDPCFPSKLKD
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: 0.06
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
Measured pKd: 3.53
PUBMED ID: 38995680

Experiment series 32

Protein A

Protein: SMCHD1
Fragment: 1139-1153
Site: PxxP_1
Sequence: FTVRPLPDEPKHLKC
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: 0.02
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 33

Protein A

Protein: SMCHD1
Fragment: 1139-1153
Site: PxxP_1
Sequence: FTVRPLPDEPKHLKC
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: 0.04
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 34

Protein A

Protein: SMCHD1
Fragment: 437-451
Site: PxxP_3
Sequence: ETYPDDPCFPSKLKD
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 35

Protein A

Protein: SMCHD1
Fragment: 1977-1991
Site: PxxP_2
Sequence: VETTDCPVPPKRMRR
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: 0.68
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: 5.7
PUBMED ID: 38995680

Experiment series 36

Protein A

Protein: SMCHD1
Fragment: 1977-1991
Site: PxxP_2
Sequence: VETTDCPVPPKRMRR
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: 0.68
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
Measured pKd: 5.14
PUBMED ID: N

Experiment series 37

Protein A

Protein: SMCHD1
Fragment: 1977-1991
Site: PxxP_2
Sequence: VETTDCPVPPKRMRR
Construct: Bio E E Ado Ado-VETTDCPVPPKRMRR-CONH2

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: R581C
Construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.31
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: 4.72
PUBMED ID: 38995680

Experiment series 38

Protein A

Protein: SMCHD1
Fragment: 1139-1153
Site: PxxP_1
Sequence: FTVRPLPDEPKHLKC
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: -0.02
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 39

Protein A

Protein: SMCHD1
Fragment: 1139-1153
Site: PxxP_1
Sequence: FTVRPLPDEPKHLKC
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
Holdup BI standard deviation: 0.05
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 40

Protein A

Protein: SMCHD1
Fragment: 437-451
Site: PxxP_3
Sequence: ETYPDDPCFPSKLKD
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
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 41

Protein A

Protein: SMCHD1
Fragment: 437-451
Site: PxxP_3
Sequence: ETYPDDPCFPSKLKD
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: 0.02
Holdup BI standard deviation: 0.02
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 42

Protein A

Protein: SMCHD1
Fragment: 1977-1991
Site: PxxP_2
Sequence: VETTDCPVPPKRMRR
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: 0.84
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
Measured pKd: 5.57
PUBMED ID: 38995680

Experiment series 43

Protein A

Protein: SMCHD1
Fragment: 1977-1991
Site: PxxP_2
Sequence: VETTDCPVPPKRMRR
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: 0.84
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
Measured pKd: 5.57
PUBMED ID: N

Experiment series 44

Protein A

Protein: SMCHD1
Fragment: 1977-1991
Site: PxxP_2
Sequence: VETTDCPVPPKRMRR
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: 0.81
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: 5.47
PUBMED ID: 38995680

Experiment series 45

Protein A

Protein: SMCHD1
Fragment: 1977-1991
Site: PxxP_2
Sequence: VETTDCPVPPKRMRR
Construct: Bio E E Ado Ado-VETTDCPVPPKRMRR-CONH2

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: T532M
Construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.48
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: 4.76
PUBMED ID: 38995680

Experiment series 46

Protein A

Protein: SMCHD1
Fragment: 1139-1153
Site: PxxP_1
Sequence: FTVRPLPDEPKHLKC
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: -0.04
Holdup BI standard deviation: 0.1
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 47

Protein A

Protein: SMCHD1
Fragment: 1139-1153
Site: PxxP_1
Sequence: FTVRPLPDEPKHLKC
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: -0.04
Holdup BI standard deviation: 0.1
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 48

Protein A

Protein: SMCHD1
Fragment: 1139-1153
Site: PxxP_1
Sequence: FTVRPLPDEPKHLKC
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
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 49

Protein A

Protein: SMCHD1
Fragment: 437-451
Site: PxxP_3
Sequence: ETYPDDPCFPSKLKD
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: 0.05
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 50

Protein A

Protein: SMCHD1
Fragment: 437-451
Site: PxxP_3
Sequence: ETYPDDPCFPSKLKD
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: 0.05
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 51

Protein A

Protein: SMCHD1
Fragment: 437-451
Site: PxxP_3
Sequence: ETYPDDPCFPSKLKD
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: 2022-10-24
Number of measurements: 2
The affinity was below the detection threshold of the assay.
PUBMED ID: 38995680

Experiment series 52

Protein A

Protein: SMCHD1
Fragment: 1977-1991
Site: PxxP_2
Sequence: VETTDCPVPPKRMRR
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: 0.77
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: 5.34
PUBMED ID: 38995680

Experiment series 53

Protein A

Protein: SMCHD1
Fragment: 1977-1991
Site: PxxP_2
Sequence: VETTDCPVPPKRMRR
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: 0.75
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
Measured pKd: 5.28
PUBMED ID: N

Experiment series 54

Protein A

Protein: SMCHD1
Fragment: 1977-1991
Site: PxxP_2
Sequence: VETTDCPVPPKRMRR
Construct: Bio E E Ado Ado-VETTDCPVPPKRMRR-CONH2

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: V566M
Construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.53
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: 4.85
PUBMED ID: 38995680

Experiment series 55

Protein A

Protein: SMCHD1
Fragment: 1139-1153
Site: PxxP_1
Sequence: FTVRPLPDEPKHLKC
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 56

Protein A

Protein: SMCHD1
Fragment: 1139-1153
Site: PxxP_1
Sequence: FTVRPLPDEPKHLKC
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: 0.02
Holdup BI standard deviation: 0.03
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 57

Protein A

Protein: SMCHD1
Fragment: 437-451
Site: PxxP_3
Sequence: ETYPDDPCFPSKLKD
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: 0.02
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 58

Protein A

Protein: SMCHD1
Fragment: 437-451
Site: PxxP_3
Sequence: ETYPDDPCFPSKLKD
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: 0.04
Holdup BI standard deviation: 0.03
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 59

Protein A

Protein: SMCHD1
Fragment: 1977-1991
Site: PxxP_2
Sequence: VETTDCPVPPKRMRR
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: 0.71
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: 5.61
PUBMED ID: 38995680

Experiment series 60

Protein A

Protein: SMCHD1
Fragment: 1977-1991
Site: PxxP_2
Sequence: VETTDCPVPPKRMRR
Construct: Bio E E Ado Ado-VETTDCPVPPKRMRR-CONH2

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: V583I
Construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.37
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: 4.7
PUBMED ID: 38995680

Experiment series 61

Protein A

Protein: SMCHD1
Fragment: 1139-1153
Site: PxxP_1
Sequence: FTVRPLPDEPKHLKC
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: 0.04
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 62

Protein A

Protein: SMCHD1
Fragment: 437-451
Site: PxxP_3
Sequence: ETYPDDPCFPSKLKD
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: -0.01
Holdup BI standard deviation: 0.06
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 63

Protein A

Protein: SMCHD1
Fragment: 1977-1991
Site: PxxP_2
Sequence: VETTDCPVPPKRMRR
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: 0.45
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: 4.71
PUBMED ID: 38995680

Experiment series 64

Protein A

Protein: SMCHD1
Fragment: 1977-1991
Site: PxxP_2
Sequence: VETTDCPVPPKRMRR
Construct: Bio E E Ado Ado-VETTDCPVPPKRMRR-CONH2

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: Y531S
Construct: his6-MBP-TEVsite-SH3
Average holdup BI: 0.14
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: 3.96
PUBMED ID: 38995680

Experiment series 65

Protein A

Protein: SMCHD1
Fragment: 1139-1153
Site: PxxP_1
Sequence: FTVRPLPDEPKHLKC
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: 0.02
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 66

Protein A

Protein: SMCHD1
Fragment: 437-451
Site: PxxP_3
Sequence: ETYPDDPCFPSKLKD
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: 0.02
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