Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving antigen-antibody binding – specific binding protein...
Reexamination Certificate
1999-08-30
2003-02-18
Kunz, Gary L. (Department: 1647)
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving antigen-antibody binding, specific binding protein...
C435S069100, C435S252300, C435S471000
Reexamination Certificate
active
06521413
ABSTRACT:
FIELD OF INVENTION
The present invention relates to nucleic acids and receptor proteins encoded thereby. Invention nucleic acids encode novel human N-methyl-D-aspartate (NMDA) receptor subunits. The invention also relates to methods for making such receptor subunits and for using the receptor proteins in assays designed to identify and characterize compounds which affect the function of such receptors, e.g., agonists and antagonists of NMDA receptors.
BACKGROUND OF THE INVENTION
The amino acid L-glutamate is a major excitatory neurotransmitter in the mammalian central nervous system. Anatomical, biochemical and electrophysiological analyses suggest that glutamatergic systems are involved in a broad array of neuronal processes, including fast excitatory synaptic transmission, regulation of neurotransmitter releases, long-term potentiation, learning and memory, developmental synaptic plasticity, hypoxic-ischemic damage and neuronal cell death, epileptiform seizures, as well as the pathogenesis of several neurodegenerative disorders. See generally, Monaghan et al., Ann. Rev. Pharmacol. Toxicol. 29:365-402 (1980). This extensive repertoire of functions, especially those related to learning, neurotoxicity and neuropathology, has stimulated recent attempts to describe and define the mechanisms through which glutamate exerts its effects.
Currently, glutamate receptor classification schemes are based on pharmacological criteria. Glutamate has been observed to mediate its effects through receptors that have been categorized into two main groups: ionotropic and metabotropic. Ionotropic glutamate receptors contain integral cation-specific, ligand-gated ion channels, whereas metabotropic glutamate receptors are G-protein-coupled receptors that transduce extracellular signals via activation of intracellular second messenger systems. Ionotropic receptors are further divided into at least two categories based on the pharmacological and functional properties of the receptors. The two main types of ionotropic receptors are N-methyl-D-aspartic acid (NMDA) and kainic acid (KA)/&agr;-amino-3-hydroxy-5-methyl-isoxazole-4-propionic acid (AMPA), formerly called the quisqualic acid, or QUIS, receptor. While the metabotropic receptors bind to some of the same ligands that bind to ionotropic glutamate receptors, the metabotropic receptors alter synaptic physiology via GTP-binding proteins and second messengers such as cyclic AMP, cyclic GMP, diacylglycerol, inositol 1,4,5-triphosphate and calcium [Gundersen et al., Proc. R. Soc. London Ser. 221:127 (1984); Sladeczek et al., Nature 317:717 (1985); Nicoletti et al., J. Neurosci. 6:1905 (1986); Sugiyama et al., Nature 325:531 (1987)].
The electrophysiological and pharmacological properties of the glutamate receptors have been studied using animal tissues and cell lines, as well as recombinantly produced non-human receptors, as the source of such receptors. The value of such studies for application to the development of human therapeutics has been limited by the availability of only non-human receptor subunits. Moreover, it is only recently that the characteristics and structure of glutamate receptors have been investigated at the molecular level. The majority of such investigation has, however, been carried out in non-human species. Because of the potential physiological and pathological significance of glutamate receptors, it would be desirable (for example, for drug screening assays) to have available human sequences (i.e., DNA, RNA, proteins) which encode representative members of the various glutamate receptor subtypes. The availability of such human sequences will also enable the investigation of receptor distribution in humans, the correlation of specific receptor modification with the occurrence of various disease states, etc.
SUMMARY OF THE INVENTION
The present invention discloses novel nucleic acids encoding NMDA receptor protein subunits and the proteins encoded thereby. In a particular embodiment the novel nucleic acids encode NMDAR1 and NMDAR2 subunits of human NMDA receptors. More specifically, the invention nucleic acids encode NMDAR1, NMDAR2A, NMDAR2B, NMDAR2C and NMDAR2D subunits that contribute to the formation of NMDA-activated cation-selective ion channels. In addition to being useful for the production of NMDA receptor subunit proteins, these nucleic acids are also useful as probes, thus enabling those skilled in the art, without undue experimentation, to identify and isolate nucleic acids encoding related receptor subunits.
Functional glutamate receptors can be assembled, in accordance with the present invention, from a plurality of NMDA receptor subunit proteins of one type (homomeric) or from combinations of subunit proteins of different types (heteromeric).
In addition to disclosing novel NMDA receptor protein subunits, the present invention also comprises methods for using such receptor subunits to identify and characterize compounds which affect the function of such receptors, e.g., agonists, antagonists, and modulators of glutamate receptor function. The invention also comprises methods for determining whether unknown protein(s) are functional as NMDA receptor subunits.
REFERENCES:
patent: 4837148 (1989-06-01), Cregg
patent: 4855231 (1989-08-01), Stroman et al.
patent: 4882279 (1989-11-01), Cregg
patent: 4929555 (1990-05-01), Cregg et al.
patent: 5024939 (1991-06-01), Gorman
patent: 5028707 (1991-07-01), Nichols et al.
patent: 5202257 (1993-04-01), Heinemann et al.
patent: 5401629 (1995-03-01), Harpold et al.
patent: 5403484 (1995-04-01), Ladner et al.
patent: 5436128 (1995-07-01), Harpold et al.
patent: 5502166 (1996-03-01), Mishina
patent: 5614509 (1997-03-01), Turski et al.
patent: 5648259 (1997-07-01), Mallet
patent: 5849895 (1998-12-01), Daggett et al.
patent: 5985586 (1999-11-01), Daggett et al.
patent: 0600278 (1994-06-01), None
patent: 0606734 (1994-07-01), None
patent: 0674003 (1995-09-01), None
patent: 2291647 (1996-01-01), None
patent: 6014783 (1994-01-01), None
patent: 9106648 (1991-05-01), None
patent: 9313423 (1993-07-01), None
patent: 9323536 (1993-11-01), None
patent: 9324629 (1993-12-01), None
patent: 9325679 (1993-12-01), None
patent: 9401094 (1994-01-01), None
patent: 9404698 (1994-03-01), None
patent: 9406428 (1994-03-01), None
patent: 9424284 (1994-04-01), None
patent: 9411501 (1994-05-01), None
patent: 9426318 (1994-11-01), None
patent: 9526401 (1995-10-01), None
Karp SJ, et al. Molecular cloning and chromosomal localization of the key subunit of the human N-Methyl-D-Aspartate receptor. J. Biol. Chem. vol. 268, pp. 3728-3733, 1993.*
Cepeda C. et al., Neurosci. Lett. 126:167-171, 1991.*
Masayuki, Human mRNA for key subunit of the N-methyl-D-aspartate receptor, DDBJ database (Jul. 20, 1993).
Abbott, NMDA receptor cloned,Trends Pharmacol. Sci. 12:449 (1991).
Abbott, NMDA receptor subunit cloned,Trends Pharmacol. Sci. 12:334 (1991).
Abe et al., Molecular characterization of a novel metabotropic glutamate receptor mGluR5 coupled to inositol phosphate/CA2+signal transduction,J. Biol. Chem. 267:13361-13368 (1992).
Albin et al., Abnormalities of striatal projection neurons and N-methyl-D-aspartate receptors in presymptomatic Huntington's Disease,N. Engl. J. Med. 322(18):1293-1298 (1990).
Anantharam et al., Combinatorial RNA splicing alters the surface charge on the NMDA receptor,FEBS Lett. 305(1):27-30 (1992).
Bahouth et al., Immunological approaches for probing receptor structure and function,Trends Pharmacol. Sci. 12:338-343 (1991).
Barnard, Will the real NMDA receptor please stand up?Trends Pharmacol. Sci. 13:11-12 (1992).
Beal, Mechanisms of excitotoxicity in neurologic diseases,FASEB J. 6:3338-3344 (1992).
Ben-Ari et al., Protein kinase C modulation of NMDA currents: an important link for LTP induction,Trends Neurosic, 15:333-339 (1992).
Black et al., N-methyl-D-aspartate- or glutamate-mediated toxicity in cultured rat cortical rat cortical neurons is antagonized by FPL 15896AR,J. Neurochem. 65:2170-2177 (1995).
Bledsoe et al., Molecular homology and DNA Hybridization, J. Mol. Evo
Daggett Lorrie P.
Ellis Steven B.
Liaw Chen Wang
Lu Chin-Chun
Giesser Joanne M.
Kohli Vineet
Kunz Gary L.
Landsman Robert S.
Merck & Co. , Inc.
LandOfFree
Human N-methyl-D-aspartate receptor subnits, nucleic acids... does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Human N-methyl-D-aspartate receptor subnits, nucleic acids..., we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Human N-methyl-D-aspartate receptor subnits, nucleic acids... will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-3121325