Interaction Details


Details of experiments between P53999 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: SUB1
Fragment: 1-127
Construct: endogenous protein (Jurkat)

Protein B

Protein: BIN1
Fragment: 513-593
Site: SH3
Construct: avi(biotin)-his6-MBP-TEVsite-SH3
Average holdup BI: 0.45
Immobilized partner concentration (10-6M): 10
P value: 7.77
Experiment method: nHU_Jurkat
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-06-29
Number of measurements: 3
Measured pKd: 4.91
PUBMED ID: 38995680
Experimental Details

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

Experiment series 2

Protein A

Protein: SUB1
Fragment: 27-41
Site: PxxP_1
Sequence: RKKQVAPEKPVKKQK
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: 0.12
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
Measured pKd: 3.9
PUBMED ID: 38995680

Experiment series 3

Protein A

Protein: SUB1
Fragment: 27-41
Site: PxxP_1
Sequence: RKKQVAPEKPVKKQK
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: 0.12
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
Measured pKd: 3.9
PUBMED ID: N

Experiment series 4

Protein A

Protein: SUB1
Fragment: 27-41
Site: PxxP_1
Sequence: RKKQVAPEKPVKKQK
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: 0.09
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
Measured pKd: 3.77
PUBMED ID: 38995680

Experiment series 5

Protein A

Protein: SUB1
Fragment: 1-127
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: 4.66
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 6

Protein A

Protein: SUB1
Fragment: 27-41
Site: PxxP_1
Sequence: RKKQVAPEKPVKKQK
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: 0.08
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 7

Protein A

Protein: SUB1
Fragment: 27-41
Site: PxxP_1
Sequence: RKKQVAPEKPVKKQK
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: 0.03
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 8

Protein A

Protein: SUB1
Fragment: 27-41
Site: PxxP_1
Sequence: RKKQVAPEKPVKKQK
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: 0.1
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
Measured pKd: 4.27
PUBMED ID: 38995680

Experiment series 9

Protein A

Protein: SUB1
Fragment: 27-41
Site: PxxP_1
Sequence: RKKQVAPEKPVKKQK
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: 0.15
Holdup BI standard deviation: 0.22
Normalized holdup BI: 0.19
Normalized holdup BI standard deviation: 0.27
Immobilized partner concentration (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022-08-04
Number of measurements: 2
Measured pKd: 4.11
PUBMED ID: N

Experiment series 10

Protein A

Protein: SUB1
Fragment: 27-41
Site: PxxP_1
Sequence: RKKQVAPEKPVKKQK
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: 0.2
Holdup BI standard deviation: 0.21
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 11

Protein A

Protein: SUB1
Fragment: 27-41
Site: PxxP_1
Sequence: RKKQVAPEKPVKKQK
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: 0.12
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
Measured pKd: 3.89
PUBMED ID: 38995680

Experiment series 12

Protein A

Protein: SUB1
Fragment: 27-41
Site: PxxP_1
Sequence: RKKQVAPEKPVKKQK
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: 0.17
Holdup BI standard deviation: 0.24
Immobilized partner concentration (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022-08-04
Number of measurements: 2
Measured pKd: 4.08
PUBMED ID: N

Experiment series 13

Protein A

Protein: SUB1
Fragment: 27-41
Site: PxxP_1
Sequence: RKKQVAPEKPVKKQK
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: 0.06
Holdup BI standard deviation: 0.09
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: 4.02
PUBMED ID: 38995680

Experiment series 14

Protein A

Protein: SUB1
Fragment: 27-41
Site: PxxP_1
Sequence: RKKQVAPEKPVKKQK
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: 0.1
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
Measured pKd: 3.82
PUBMED ID: 38995680

Experiment series 15

Protein A

Protein: SUB1
Fragment: 27-41
Site: PxxP_1
Sequence: RKKQVAPEKPVKKQK
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: 0.07
Holdup BI standard deviation: 0.09
Immobilized partner concentration (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022-06-29
Number of measurements: 2
Measured pKd: 3.61
PUBMED ID: 38995680

Experiment series 16

Protein A

Protein: SUB1
Fragment: 27-41
Site: PxxP_1
Sequence: RKKQVAPEKPVKKQK
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: 0.07
Holdup BI standard deviation: 0.09
Immobilized partner concentration (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022-06-29
Number of measurements: 2
Measured pKd: 3.61
PUBMED ID: N

Experiment series 17

Protein A

Protein: SUB1
Fragment: 27-41
Site: PxxP_1
Sequence: RKKQVAPEKPVKKQK
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: 0.19
Holdup BI standard deviation: 0.2
Immobilized partner concentration (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022-08-04
Number of measurements: 2
Measured pKd: 4.13
PUBMED ID: N

Experiment series 18

Protein A

Protein: SUB1
Fragment: 27-41
Site: PxxP_1
Sequence: RKKQVAPEKPVKKQK
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: 0.06
Holdup BI standard deviation: 0.09
Immobilized partner concentration (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022-10-24
Number of measurements: 2
Measured pKd: 3.59
PUBMED ID: 38995680

Experiment series 19

Protein A

Protein: SUB1
Fragment: 27-41
Site: PxxP_1
Sequence: RKKQVAPEKPVKKQK
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: 0.06
Holdup BI standard deviation: 0.09
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: 3.82
PUBMED ID: 38995680

Experiment series 20

Protein A

Protein: SUB1
Fragment: 27-41
Site: PxxP_1
Sequence: RKKQVAPEKPVKKQK
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: 0.11
Holdup BI standard deviation: 0.09
Immobilized partner concentration (10-6M): 18
Experiment method: HU (DAPF)
Date of measurement: 2022-10-24
Number of measurements: 2
Measured pKd: 3.85
PUBMED ID: 38995680