Alzheimer's disease secretase

Details Alzheimer's disease secretase Alzheimer's disease secretase

2001-09-17

2004-12-28

Kemmerer, Elizabeth (Department: 1647)

Chemistry: molecular biology and microbiology

Micro-organism, per se ; compositions thereof; proces of...

Bacteria or actinomycetales; media therefor

C530S300000, C530S350000, C536S023100, C536S023500, C435S254110, C435S320100, C435S325000, C435S352000, C435S363000, C435S366000, C435S367000

Reexamination Certificate

active

06835565

ABSTRACT:

FIELD OF THE INVENTION
The present invention related to the field of Alzheimer's Disease. APP, amyloid beta peptide, and human aspartyl proteases as well as a method for the identification of agents that modulate the activity of these polypeptides.
BACKGROUND OF THE INVENTION
Alzheimer's disease (AD) causes progressive dementia with consequent formation of amyloid plaques, neurofibrillary tangles, gliosis and neuronal loss. The disease occurs in both genetic and sporadic forms whose clinical course and pathological features are quite similar. Three genes have been discovered to date which when mutated cause an autosomal dominant form of Alzheimer's disease. These encode the amyloid protein precursor (APP) and two related proteins, presenilin-1 (PS1) and presenilin-2 (PS2), which as their names suggest are both structurally and functionally related. Mutations in any of the three enhance proteolytic processing of APP via an intracellular pathway that produces amyloid beta peptide or the A&bgr; peptide (or sometimes here as Abeta), a 40-42 amino acid long peptide that is the primary component of amyloid plaque in AD. Dysregulation of intracellular pathways for proteolytic processing may be central to the pathophysiology of AD. In the case of plaque formation, mutations in APP, PS1 or PS2 consistently alter the proteolytic processing of APP so as to enhance formation of A&bgr; 1-42, a form of the A&bgr; peptide which seems to be particularly amyloidogenic, and thus very important in AD. Different forms of APP range in size from 695-770 amino acids, localize to the cell surface, and have a single C-terminal transmembrane domain. The Abeta peptide is derived from a region of APP adjacent to and containing a portion of the transmembrane domain. Normally, processing of APP at the &agr;-secretase site cleaves the midregion of the A&bgr; sequence adjacent to the membrane and releases the soluble, extracellular domain of APP from the cell surface. This &agr;-secretase APP processing, creates soluble APP-&agr;, and it is normal and not thought to contribute to AD.
Pathological processing of APP at the &bgr;- and &ggr;-secretase sites produces a very different result than procesing at the &agr; site. Sequential processing at the &bgr;- and &ggr;-secretase sites releases the A&bgr; peptide, a peptide possibly very important in AD pathogenesis. Processing at the &bgr;- and &ggr;-secretase sites can occur in both the endoplasmic reticulum (in neurons) and in the endosomal/lysosomal pathway after reinternalization of cell surface APP (in all cells). Despite intense efforts, for 10 years or more, to identify the enzymes responsible for processing APP at the &bgr; and &ggr; sites, to produce the &bgr; peptide, those proteases remained unknown until this disclosure. Here, for the first time, we report the identification and characterization of the &bgr; secretase enzyme. We disclose some known and some novel human aspartic proteases that can act as &bgr;-secretase proteases and, for the first time, we explain the role these proteases have in AD. We describe regions in the proteases critical for their unique function and for the first time characterize their substrate. This is the first description of expressed isolated purified active protein of this type, assays that use the protein, in addition to the identification and creation of useful cell lines and inhibitors.
SUMMARY OF THE INVENTION
Here we disclose a number of variants of the asp2 gene and peptide.
The invention provides for an isolated or purified nucleic acid polynucleotide that codes for a protease capable of cleaving the beta (&bgr;) secretase cleavage site of APP that contains two or more sets of special nucleic acids, where the special nucleic acids are separated by nucleic acids that code for about 100 to 300 amino acid positions, where the amino acids in those positions may be any amino acids, where the first set of special nucleic acids consists of the nucleic acids that code for the peptide DTG, where the first nucleic acid of the first special set of nucleic acids is, the first special nucleic acid, and where the second set of nucleic acids code for either the peptide DSG or DTG, where the last nucleic acid of the second set of nucleic acids is the last special nucleic acid, with the proviso that the nucleic acids disclosed in SEQ ID NO. 1 and SEQ. ID NO. 5 are not included. In one embodiment, the two sets of nucleic acids are separated by nucleic acids that code for about 125 to 222 amino acid positions, which may be any amino acids. In a particular embodiment, the two sets of special nucleic acids are separated by nucleic acids that code for about 150 to 172 amino acid positions, which may be any amino acids. In a more particular embodiment, the two sets are separated by nucleic acids that code for about 172 amino acid positions, which may be any amino acids. An exemplary polynucleotide comprises the nucleic acid described in SEQ. ID. NO. 3. In another embodiment, the two sets of nucleic acids are separated by nucleic acids that code for about 150 to 196 amino acid positions. In another embodiment, the two sets of nucleotides are separated by nucleic acids that code for about 196 amino acids. An exemplary polynucleotide comprises the two sets of nucleic acids separated by the same nucleic acid sequences that separate the same set of special nucleic acids in SEQ. ID. NO. 5. In a particular embodiment, the two sets of nucleic acids are separated by nucleic acids that code for about 150 to 190, amino acid. In another embodiment, the two sets of nucleotides are separated by nucleic acids that code for about 190 amino acids. In a more particular embodiment, the two sets of nucleotides are separated by the same nucleic acid sequences that separate the same set of special nucleotides in SEQ. ID. NO. 1. In one embodiment, the first nucleic acid of the first special set of amino acids, that is, the first special nucleic acid, is operably linked to any codon where the nucleic acids of that codon codes for any peptide comprising from 1 to 10,000 amino acid. In one variation, the first special nucleic acid is operably linked to nucleic acid polymers that code for any peptide selected from the group consisting of: any any reporter proteins or proteins which facilitate purification. In another variation, the first special nucleic acid is operably linked to nucleic acid polymers that code for any peptide selected from the group consisting of: immunoglobin-heavy chain, maltose binding protein, glutathion S transfection, Green Fluorescent protein, and ubiquitin. In another embodiment, the last nucleic acid of the second set of special amino acids, that is, the last special nucleic acid, is operably linked to nucleic acid polymers that code for any peptide comprising any amino acids from 1 to 10,000 amino acids. In one variation, the last special nucleic acid is operably linked to any codon linked to nucleic acid polymers that code for any peptide selected from the group consisting of: any reporter proteins or proteins which facilitate purification. In another embodiment, the first special nucleic acid is operably linked to nucleic acid polymers that code for any peptide selected from the group consisting of: immunoglobin-heavy chain, maltose binding protein, glutathion S transfection, Green Fluorescent protein, and ubiquitin.
The invention provides for an isolated or purified nucleic acid polynucleotide that codes for a protease capable of cleaving the beta secretase cleavage site of APP that contains two or more sets of special nucleic acids, where the special nucleic acids are separated by nucleic acids that code for about 100 to 300 amino acid positions, where the amino acids in those positions may be any amino acids, where the first set of special nucleic acids consists of the nucleic acids that code for DTG, where the first nucleic acid of the first special set of nucleic acids is, the first special nucleic acid, and where the second set of nucleic acids code for either DSG or DTG, where the last nucleic acid of the sec

Affiliated with

Also associated with

Rating

Alzheimer's disease secretase does not yet have a rating. At this time, there are no reviews or comments for this patent.

If you have personal experience with Alzheimer's disease secretase, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Alzheimer's disease secretase will most certainly appreciate the feedback.

Rate now

Comments { 0 }

Profile ID: LFUS-PAI-O-3281847