Chemistry: natural resins or derivatives; peptides or proteins; – Proteins – i.e. – more than 100 amino acid residues – Blood proteins or globulins – e.g. – proteoglycans – platelet...
Reexamination Certificate
1998-07-21
2001-04-03
Kunz, Gary L. (Department: 1647)
Chemistry: natural resins or derivatives; peptides or proteins;
Proteins, i.e., more than 100 amino acid residues
Blood proteins or globulins, e.g., proteoglycans, platelet...
C530S387900, C424S141100, C424S139100
Reexamination Certificate
active
06211340
ABSTRACT:
BACKGROUND OF THE INVENTION
The dystrophin-glycoprotein complex (DGC) in skeletal muscle is a complex of sarcolemmal proteins and glycoproteins. It is composed of dystrophin, a cytoskeletal actin-binding protein; the syntrophins, a 59 kDa triplet of intracellular proteins that bind the C-terminal domain of dystrophin; &agr;-dystroglycan, a 156 kDa extracellular proteoglycan which binds the G domain of laminin; &bgr;-dystroglycan, a 43 kDa transmembrane glycoprotein which binds the cysteine-rich region of dystrophin, &agr;-, &bgr;-, and &ggr;-sarcoglycan, transmembrane glycoproteins of 50, 43, and 35 kDa respectively; and a 25 kDa transmembrane protein. Recent experiments have demonstrated the existence of two complexes within the DGC: the dystroglycan complex, composed of &agr;- and &bgr;-dystroglycan, and the sarcoglycan complex, consisting of &agr;-, &bgr;-, and &ggr;-sarcoglycan.
Defects in DGC components lead to muscle fiber necrosis, the major pathological event in muscular dystrophies. In Duchenne muscular dystrophy (DMD), mutations in the dystrophin gene cause the loss of dystrophin and a reduction of the dystrophin-associated proteins. One form of congenital muscular dystrophy (CMD) has recently been characterized as being caused by mutations in the laminin &agr;2-chain gene. Limb-girdle muscular dystrophy (LGMD) represents a clinically and genetically heterogeneous class of disorders. They are inherited as either autosomal dominant or recessive traits. An autosomal dominant form, LGMD1A, was mapped to 5q31-q33 (Speer, M. C. et al.,
Am. J. Hum. Genet
. 50:1211, 1992; Yamaoka, L. Y. et al.,
Neuromusc. Disord
. 4:471, 1994), while six genes involved in the autosomal recessive forms were mapped to 15q15.1 (LGMD2A)(Beckmann, J. S. et al.,
C. R. Acad. Sci. Paris
312:141, 1991), 2p16-pl3 (LGMD2B)(Bashir, R. et al.,
Hum. Mol. Genet
. 3:455, 1994), 13q12 (LGMD2C) (Ben Othmane, K. et al.,
Nature Genet
. 2:315, 1992; Azibi, K. et al.,
Hum. Mol. Genet
. 2:1423, 1993), 17q12-q21.33 (LGMD2D)(Roberds, S. L. et al.,
Cell
78:625, 1994; McNally, E. M., et. al.,
Proc. Nat. Acad. Sci. U.S.A
. 91:9690, 1994), 4q12 (LG1MD2E) (Lim, L. E., et. al.,
Nat. Genet
. 11:257, 1994; Bönnemann, C. G. et al.
Nat. Genet
. 11:266, 1995), and most recently to 5q33-q34 (LGMD2F)(Passos-Bueno, M. R., et. al.,
Hum. Mol. Genet
. 5:815, 1996). Patients with LGMD2C, 2D and 2E have a deficiency of components of the sarcoglycan complex resulting from mutations in the genes encoding &ggr;-, &agr;-, and &bgr;-sarcoglycan respectively. The gene responsible for LGMD2A has been identified as the muscle-specific calpain, whereas the genes responsible for LGMD1A, 2B and 2F are still unknown.
SUMMARY OF THE INVENTION
In one aspect, the present invention relates to a substantially pure nucleic acid sequence encoding a mammalian 35 kDa non-dystrophin component (&dgr;-sarcoglycan) of the dystrophin-glycoprotein complex. The substantially pure nucleic acid sequence is characterized by the ability to hybridize to the DNA sequence of SEQ ID NO:1, or the complement thereof, under stringent hybridization conditions. The substantially pure nucleic acid molecule of the present invention can also be characterized as encoding the amino acid sequence shown in SEQ ID NO:2, or equivalents of said amino acid sequence. The invention also encompasses DNA expression constructs incorporating the substantially pure nucleic acid sequence encoding &dgr;-sarcoglycan, and cells (prokaryotic and eukaryotic) which harbor such an expression construct. Such compositions are useful, for example, in the production of highly pure immunogen for use in stimulating the production of polyclonal and monoclonal antibodies.
In another aspect, the present invention relates to immunogenic peptides (or equivalents thereof) which, when used to immunize a mammal, stimulate the production of antibodies which bind specifically to the &dgr;-sarcoglycan. Such peptides are useful, for example, in the production of highly pure immunogen for use in stimulating the production of polyclonal and monoclonal antibodies.
Another aspect of the present invention relates to direct sequencing methods for the determination of mutations responsible for disorders such as autosomal recessive limb-girdle muscular dystrophy. Based on the information obtained through direct sequencing, nucleic acid probes can be designed which hybridize specifically to a mutant form of &dgr;-sarcoglycan, or the complement thereof, but not to the DNA of the wild-type form of the gene (or the complement thereof), under stringent hybridization conditions. Such probes are useful, for example, in connection with the diagnosis of autosomal recessive limb-girdle muscular dystrophy.
REFERENCES:
Geysen et al. J. Molecular Recognition 1: 32-40, 1988.*
Nigro et al. Human Molecular Genetics 5: 1179-1186, 1986.*
Duclos et al. Amer. J. Human Genetics 59(4, Suppl.):A38, 1996.*
Straub et al. Amer. J. Human Genetics 59(4, Suppl.):A286, 1996.*
Roberds et al.,Cell 78: 625 (1994).
Noguchi et al.,Science 270: 819 (1995).
Jung et al.,FEBS Lett. 381: 15 (1996).
Passos-Bueno et al.,Hum. Mol. Genet. 5: 815 (1996).
Bento Soares et al.,Proc. Natl. Acad. Sci. U.S.A. 91: 9228 (1994).
Ohlendieck and Campbell,J. Cell. Biol. 115: 1685 (1991).
Ohlendieck et al.,J. Cell. Biol. 112: 135 (1991).
Yamamoto et al.,J. Biochem. 114: 132 (1993).
Campbell Kevin P.
Duclos Franck
Jung Daniel
McPherson John
Straub Volker
Farrell Kevin M.
Hayes Robert C.
Kunz Gary L.
University of Iowa Research Foundation
LandOfFree
&dgr;-sarcoglycan antibodies does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with &dgr;-sarcoglycan antibodies, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and &dgr;-sarcoglycan antibodies will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-2550253