Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Recombinant dna technique included in method of making a...
Reexamination Certificate
1999-11-19
2001-05-08
Park, Hankyel T. (Department: 1648)
Chemistry: molecular biology and microbiology
Micro-organism, tissue cell culture or enzyme using process...
Recombinant dna technique included in method of making a...
C435S334000, C530S326000, C530S387900, C530S388220
Reexamination Certificate
active
06228616
ABSTRACT:
FIELD OF THE INVENTION
This invention relates to nucleic acid and amino acid sequences of a novel human anion channel and to the use of these sequences in the diagnosis, prevention, and treatment of cancer and developmental disorders.
BACKGROUND OF THE INVENTION
Chloride channels are found in the plasma membranes of virtually every cell in the body. Chloride channels mediate a variety of cellular functions including regulation of membrane potentials and absorption and secretion of ion across epithelial membranes. When present in intracellular membranes of the Golgi apparatus and endocytic vesicles, chloride channels also regulate organelle pH (cf. Greger, R. (1988) Annu. Rev. Physiol. 50:111-122). Electrophysiological and pharmacological measurements including ion conductance, current-voltage relationships, and sensitivity to modulators suggest that different chloride channels exist in muscles, neurons, fibroblasts, epithelial cells, and lymphocytes.
Several chloride channels have been cloned from mammalian tissues and cell lines. The sequences of these proteins are diverse and include a nicotinic acetylcholine receptor homolog, the cystic fibrosis transmembrane conductance regulator, and the highly acidic p64 protein with no significant homology to other proteins (Bernard, E. A. et al. (1987) Trends Neurosci. 16:502-509; Riordan, J. R. et al. (1989) Science 245:1066-1073; Landry, D. et al. (1993) J. Biol. Chem. 268:14948-14955). Many of the channels have sites for phosphorylation by one or more protein kinases including protein kinase A, protein kinase C, tyrosine kinase, and casein kinase II which regulate chloride channel activity in epithelial cells.
The p64 protein was originally identified in bovine kidney cortex membranes by its affinity for indanyloxyacetic acid, a known inhibitor of epithelial chloride channels (Landry et al. (1989) Science 244:1469-1472). Antibodies raised against the isolated p64 protein can deplete solubilized kidney membranes of all detectable chloride channel activity. Thus, p64 is likely to be a functional component of the kidney chloride channel (Redhead, R. C. et al. (1992) Proc. Natl. Acad. Sci. 89:3716-3720).
Northern blot analyses using the p64 clone as a probe detect related mRNAs, ranging in size from ~2 kb to ~6.5 kb, in bovine kidney cortex, kidney medulla, liver, adrenal glands, brain, skeletal muscle, and heart. Most of these tissues have multiple transcripts that are capable of hybridizing to this probe. The diversity and relative abundance of these transcripts is tissue-specific, and this suggests that the p64 transcripts are alternatively spliced and/or that a family of related genes are expressed (Landry et al., supra).
The sequence of p64 predicts an acidic, integral membrane protein which spans the membrane at least twice and has the amino terminus on the cytoplasmic side. The protein has potential sites for phosphorylation by protein kinase C, tyrosine kinase, and casein kinase II, and a single site for N-linked glycosylation at Asp
235
.
The discovery of polynucleotides encoding a novel human anion channel, and the molecules themselves, provides a means to investigate the regulation of membrane potentials, intracellular pH, cell volume, signal transduction, and transepithelial ion transport in tissues containing absorptive or secretory epithelia under normal and disease conditions. Discovery of a novel anion channel satisfies a need in the art by providing new compositions useful in diagnosing and treating cancer and developmental disorders.
SUMMARY OF THE INVENTION
The present invention features a novel human anion channel protein hereinafter designated NANCH and characterized as having similarity to bovine p64 chloride channel and human P64CLCP.
Accordingly, the invention features a substantially purified NANCH having the amino acid sequence shown in SEQ ID NO: 1.
One aspect of the invention features isolated and substantially purified polynucleotides that encode NANCH. In a particular aspect, the polynucleotide is the nucleotide sequence of SEQ ID NO:2.
The invention also relates to a polynucleotide sequence comprising the complement of SEQ ID NO:2 or variants thereof. In addition, the invention features polynucleotide sequences which hybridize under stringent conditions to SEQ ID NO:2.
The invention additionally features nucleic acid sequences encoding polypeptides, oligonucleotides, peptide nucleic acids (PNA), fragments, portions or antisense molecules thereof, and expression vectors and host cells comprising polynucleotides that encode NANCH. The present invention also features antibodies which bind specifically to NANCH, and pharmaceutical compositions comprising substantially purified NANCH. The invention also features the use of agonists and antagonists of NANCH. The invention also features methods for treating disorders which are associated with NANCH, and for detecting a polynucleotide which encodes NANCH.
REFERENCES:
Greger, R., “Chloride Transport in Thick Ascending Limb, Distal Convolution, and Collecting Duct”Annu. Rev. Physiol.50:111-122 (1988).
Bernard, E.A. et al., “Molecular biology of the GAVAAreceptor: the receptor/channel superfamily”Trends Neurosci.16:502-509 (1987).
Riordan, JR et al., “Identification of the Cystic Fibrosis Geme: Cloning and Characterization of Complementary DNA”Science245:1066-1073 (1989).
Landry, D et al., “Molecular Cloning and Characterization of p64, a Chloride Channel Protein from Kidney Microsomes”J. Biol. Chem.268:14948-14955 (1993).
Landry, DW et al., “Purification and Reconstitution of Chloride Channels from Kidney and Trachea”Science244:1469-1472 (1989).
Redhead, CR et al., “A ubiquitous 64-kDa protein is a component of a chloride channel of plasma and intracellular membranes”Proc. Natl. Acad. Sci.89:3716-3720 (1992).
Borsani, G, (GI 895845) GenBank Sequence Database (Accession 895845), National Center for Biotechnology Information, National Library of Medicine, Bethesda, Maryland, 20894 (Jan. 24, 1997).
Borsani, G, (GI 895844) GenBank Sequence Database (Accesson X87689), National Center for Biotechnology Information, National Library of Medicine, Bethesda, Maryland, 20894 (Jan. 24, 1997).
Howell, S et al., “Identification and characterisation of a homologue of p64 in rat tissues”FEBS Letter390:207-210 (1996).
Bandman Olga
Goli Surya K.
Incyte Genomics Inc.
Incyte Genomics, Inc.
Park Hankyel T.
LandOfFree
Human anion channel 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 anion channel, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Human anion channel will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-2443957