A-beta-peptide screening
AssigneeAventis Pharma Deutschland GmbH
InventorsEdmund Hoppe, Gisela Peraus, Jonathan Rothblatt, Ekkehard Leberer, Luc Mercken, Sylvie Dreisler
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Abstract
The present invention relates to an improved process for determining γ-secretase activity and for detecting γ-secretase, a subunit protein of γ-secretase, or a γ-secretase-like proteinase; particular embodiments of the process relate on the one hand to processes for the identification of a γ-secretase or of a cDNA which codes for a γ-secretase, a subunit protein of γ-secretase, or a γ-secretase-like proteinase, and on the other hand to processes for the identification of substances which can inhibit the activity of a γ-secretase, a subunit protein of γ-secretase, or a γ-secretase-like proteinase.
Claims48
A method for detecting the activity of γ-secretase, the method comprising the steps of:
- A. providing a transgene encoding a fusion protein comprising:
- a) a first nucleotide sequence encoding an amino acid sequence comprising GAIIGLMVGGVVIATVIVITLVML (SEQ ID NO. 1);
- b) a second nucleotide sequence, at the 5′ end of the first nucleotide sequence, encoding an amino acid sequence comprising a signal peptide;
- and
- c) a promoter;
- d) wherein the fusion protein does not comprise, with the exception of SEQ ID NO. 1, one or more sequences consisting of a protease cleavage site;
- and
- e) wherein the fusion protein does not comprise, with the exception of SEQ ID NO. 1, one or more sequences consisting of a signal for endo- or exocytosis.
- B. expressing the fusion protein in a cell that contains γ-secretase;
- C. detecting a polypeptide selected from the group consisting of a polypeptide that comprises GAIIGLMVGGVV (SEQ ID NO. 2) and a polypeptide that comprises VIVITLVML (SEQ ID NO. 3)
A transgene comprising
- A. a first nucleotide sequence encoding an amino acid sequence comprising GAIIGLMVGGVVIATVIVITLVML (SEQ ID NO. 1);
- B. a second nucleotide sequence, at the 5′ end of the first nucleotide sequence, encoding an amino acid sequence comprising a signal peptide;
- C. a promoter;
- and
- D. at least one additional nucleotide sequence, at the 3′ end of the first nucleotide sequence, encoding an amino acid sequence comprising a DNA-binding domain and a transcription-activating domain.
An assay comprising the steps of
- A. providing a transgenic cell, the cell comprising
- a) a first nucleotide sequence encoding an amino acid sequence comprising GAIIGLMVGGVVIATVIVITLVML (SEQ ID NO. 1);
- b) a second nucleotide sequence, at the 5′ end of the first nucleotide sequence, encoding an amino acid sequence comprising a signal peptide;
- and
- c) a promoter;
- B. Incubating the organism or cell with a substance;
- C. Determining whether a protein comprising the amino acid sequence of SEQ ID NO. 3 is formed.
Dependencies are inferred from the claim text.