Interaction Details


Details of experiments between O00499 and P30291
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: WEE1
Fragment: 1-646
Construct: endogenous protein (Jurkat)
Average holdup BI: 0.76
Immobilized partner concentration (10-6M): 10
P value: 2.81
Experiment method: nHU_Jurkat
Details of affinity fitting: hyperbolic binding equation
Date of measurement: 2022-06-29
Number of measurements: 3
Measured pKd: 5.5
PUBMED ID: 38995680
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: his6-MBP-TEVsite-SH3

Protein B

Protein: WEE1
Fragment: 3-17
Site: PxxP_1
Sequence: FLSRQQPPPPRRAGA
Construct: Bio Ado Ado-FLSRQQPPPPRRAGA-CONH2
Average holdup BI: 0.11
Holdup BI standard deviation: 0.01
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: 2
Measured pKd: 3.86
PUBMED ID: 38995680

Experiment series 3

Protein A

Protein: BIN1
Fragment: 1-593
Construct: endogenous proteins (Jurkat)

Protein B

Protein: WEE1
Fragment: 228-247
Construct: Biotin-Ahx-RQTPQVNINPFTPDSLLLHS-CONH2
Average holdup BI: 0.04
Immobilized partner concentration (10-6M): 10
P value: 0.02
Experiment method: nHU_Jurkat
Details of affinity fitting: hyperbolic equation
Date of measurement: 2024-04-11
Number of measurements: 6
The affinity was below the detection threshold of the assay.
Experimental Details

Batch = Helsinki_batch7; Ncontrols = 24; Nsamples = 6; Npreys = 6872; threshold curvature: 2.12

Experiment series 4

Protein A

Protein: BIN1
Fragment: 513-593
Site: SH3
Construct: his6-MBP-TEVsite-SH3

Protein B

Protein: WEE1
Fragment: 55-69
Site: PxxP_2
Sequence: FQEPDSPLPPARSPT
Construct: Bio Ado Ado-FQEPDSPLPPARSPT-CONH2
Average holdup BI: 0.02
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
The affinity was below the detection threshold of the assay.
PUBMED ID: 38995680

Experiment series 5

Protein A

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: D537V
Construct: his6-MBP-TEVsite-SH3

Protein B

Protein: WEE1
Fragment: 3-17
Site: PxxP_1
Sequence: FLSRQQPPPPRRAGA
Construct: Bio Ado Ado-FLSRQQPPPPRRAGA-CONH2
Average holdup BI: -0
Holdup BI standard deviation: 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: 2
The affinity was below the detection threshold of the assay.
PUBMED ID: 38995680

Experiment series 6

Protein A

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: D537V
Construct: his6-MBP-TEVsite-SH3

Protein B

Protein: WEE1
Fragment: 55-69
Site: PxxP_2
Sequence: FQEPDSPLPPARSPT
Construct: Bio Ado Ado-FQEPDSPLPPARSPT-CONH2
Average holdup BI: 0.01
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 7

Protein A

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: F584S
Construct: his6-MBP-TEVsite-SH3

Protein B

Protein: WEE1
Fragment: 3-17
Site: PxxP_1
Sequence: FLSRQQPPPPRRAGA
Construct: Bio Ado Ado-FLSRQQPPPPRRAGA-CONH2
Average holdup BI: -0.02
Holdup BI standard deviation: 0.05
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: 2
The affinity was below the detection threshold of the assay.
PUBMED ID: 38995680

Experiment series 8

Protein A

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: F584S
Construct: his6-MBP-TEVsite-SH3

Protein B

Protein: WEE1
Fragment: 55-69
Site: PxxP_2
Sequence: FQEPDSPLPPARSPT
Construct: Bio Ado Ado-FQEPDSPLPPARSPT-CONH2
Average holdup BI: -0.01
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 9

Protein A

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: P551L
Construct: his6-MBP-TEVsite-SH3

Protein B

Protein: WEE1
Fragment: 3-17
Site: PxxP_1
Sequence: FLSRQQPPPPRRAGA
Construct: Bio Ado Ado-FLSRQQPPPPRRAGA-CONH2
Average holdup BI: 0.03
Holdup BI standard deviation: 0.02
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: 2
The affinity was below the detection threshold of the assay.
PUBMED ID: 38995680

Experiment series 10

Protein A

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: P551L
Construct: his6-MBP-TEVsite-SH3

Protein B

Protein: WEE1
Fragment: 55-69
Site: PxxP_2
Sequence: FQEPDSPLPPARSPT
Construct: Bio Ado Ado-FQEPDSPLPPARSPT-CONH2
Average holdup BI: 0
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
The affinity was below the detection threshold of the assay.
PUBMED ID: 38995680

Experiment series 11

Protein A

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: Q540H
Construct: his6-MBP-TEVsite-SH3

Protein B

Protein: WEE1
Fragment: 3-17
Site: PxxP_1
Sequence: FLSRQQPPPPRRAGA
Construct: Bio Ado Ado-FLSRQQPPPPRRAGA-CONH2
Average holdup BI: 0.17
Holdup BI standard deviation: 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: 2
Measured pKd: 4.07
PUBMED ID: 38995680

Experiment series 12

Protein A

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: Q540H
Construct: his6-MBP-TEVsite-SH3

Protein B

Protein: WEE1
Fragment: 55-69
Site: PxxP_2
Sequence: FQEPDSPLPPARSPT
Construct: Bio Ado Ado-FQEPDSPLPPARSPT-CONH2
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 13

Protein A

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: R581C
Construct: his6-MBP-TEVsite-SH3

Protein B

Protein: WEE1
Fragment: 3-17
Site: PxxP_1
Sequence: FLSRQQPPPPRRAGA
Construct: Bio Ado Ado-FLSRQQPPPPRRAGA-CONH2
Average holdup BI: 0.01
Holdup BI standard deviation: 0.03
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: 2
The affinity was below the detection threshold of the assay.
PUBMED ID: 38995680

Experiment series 14

Protein A

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: R581C
Construct: his6-MBP-TEVsite-SH3

Protein B

Protein: WEE1
Fragment: 55-69
Site: PxxP_2
Sequence: FQEPDSPLPPARSPT
Construct: Bio Ado Ado-FQEPDSPLPPARSPT-CONH2
Average holdup BI: 0.03
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
The affinity was below the detection threshold of the assay.
PUBMED ID: 38995680

Experiment series 15

Protein A

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: T532M
Construct: his6-MBP-TEVsite-SH3

Protein B

Protein: WEE1
Fragment: 3-17
Site: PxxP_1
Sequence: FLSRQQPPPPRRAGA
Construct: Bio Ado Ado-FLSRQQPPPPRRAGA-CONH2
Average holdup BI: 0.13
Holdup BI standard deviation: 0.01
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: 2
Measured pKd: 3.94
PUBMED ID: 38995680

Experiment series 16

Protein A

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: T532M
Construct: his6-MBP-TEVsite-SH3

Protein B

Protein: WEE1
Fragment: 55-69
Site: PxxP_2
Sequence: FQEPDSPLPPARSPT
Construct: Bio Ado Ado-FQEPDSPLPPARSPT-CONH2
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: 2023-01-16
Number of measurements: 1
The affinity was below the detection threshold of the assay.
PUBMED ID: 38995680

Experiment series 17

Protein A

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: V566M
Construct: his6-MBP-TEVsite-SH3

Protein B

Protein: WEE1
Fragment: 3-17
Site: PxxP_1
Sequence: FLSRQQPPPPRRAGA
Construct: Bio Ado Ado-FLSRQQPPPPRRAGA-CONH2
Average holdup BI: 0.12
Holdup BI standard deviation: 0.01
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: 2
Measured pKd: 3.89
PUBMED ID: 38995680

Experiment series 18

Protein A

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: V566M
Construct: his6-MBP-TEVsite-SH3

Protein B

Protein: WEE1
Fragment: 55-69
Site: PxxP_2
Sequence: FQEPDSPLPPARSPT
Construct: Bio Ado Ado-FQEPDSPLPPARSPT-CONH2
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 19

Protein A

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: V583I
Construct: his6-MBP-TEVsite-SH3

Protein B

Protein: WEE1
Fragment: 3-17
Site: PxxP_1
Sequence: FLSRQQPPPPRRAGA
Construct: Bio Ado Ado-FLSRQQPPPPRRAGA-CONH2
Average holdup BI: 0
Holdup BI standard deviation: 0.02
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: 2
The affinity was below the detection threshold of the assay.
PUBMED ID: 38995680

Experiment series 20

Protein A

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: V583I
Construct: his6-MBP-TEVsite-SH3

Protein B

Protein: WEE1
Fragment: 55-69
Site: PxxP_2
Sequence: FQEPDSPLPPARSPT
Construct: Bio Ado Ado-FQEPDSPLPPARSPT-CONH2
Average holdup BI: 0
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
The affinity was below the detection threshold of the assay.
PUBMED ID: 38995680

Experiment series 21

Protein A

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: Y531S
Construct: his6-MBP-TEVsite-SH3

Protein B

Protein: WEE1
Fragment: 3-17
Site: PxxP_1
Sequence: FLSRQQPPPPRRAGA
Construct: Bio Ado Ado-FLSRQQPPPPRRAGA-CONH2
Average holdup BI: 0.02
Holdup BI standard deviation: 0
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: 2
The affinity was below the detection threshold of the assay.
PUBMED ID: 38995680

Experiment series 22

Protein A

Protein: BIN1
Fragment: 513-593
Site: SH3
Modification: Y531S
Construct: his6-MBP-TEVsite-SH3

Protein B

Protein: WEE1
Fragment: 55-69
Site: PxxP_2
Sequence: FQEPDSPLPPARSPT
Construct: Bio Ado Ado-FQEPDSPLPPARSPT-CONH2
Average holdup BI: 0.01
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