Calpain and DNA encoding the same

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

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C530S326000, C530S350000

Reexamination Certificate

active

06582932

ABSTRACT:

FIELD OF THE INVENTION
The present invention relates to newly identified calpain, Rt88 protein, which has been of the retina in eye tissues, and a DNA encoding it.
BACKGROUND OF THE INVENTION
Calpain is present, in particular, in the cytoplasm of animal cells and is a cysteine protease which is activated by calcium. Several molecular species have been known in calpain. For analyzing the structure, their cDNA's have been cloned and, at present, the presence of &mgr;-and m-calpain which are generally expressed in various tissues, as well as tissue-specific calpain such as, for example, p94 which is specifically expressed in a skeleton muscle is revealed [Seikagaku (Biochemistry), Vol. 65, No. 7, pp. 537-552 (1993); Jikken Igaku (Experimental Medicine), Vol. 13, No. 9, pp. 35-42 (1995)].
Although details of physiological functions of calpain are not yet elucidated, calpain has been considered to have functions of a calcium receptor in cells and to be concerned in, for example, signal transduction, control of transcription, propagation and differentiation of cells, and the like.
Recently, it has been reported that a mutant gene of calpain p94 specifically expressed in a skeleton muscle is one of causative genes of a kind of dystrophy, myodystropy, which is known to be a disease wherein differentiated cells fall into spontaneous degeneration or atrophy without any anticipation disorder such as inflammation or injury (Isabelle Richard et al., Cell, 81, 28-40 (1995)). In addition, it has been found that p94 protein is decreased in myodystrophy (Melissa J. Spencer et al., Journal of the Neurological Science, 146, 173-178 (1997)).
On the other hand, in general, retinal degenerative diseases are divided into dystrophy and other degenerative diseases. Dystrophy is hereditary and, in many cases, the prognosis of vision is pessimistic. Then, dystrophy is of importance from clinical viewpoint (Yoshihiro Hotta, “The Cause of Retinal Degeneration” in Atarashii Ganka (Journal of the Eye), 13 (7): 993-1001, 1996). In particular, at present, pigmentary retinal degeneration is designated as an objective disease in the Ministry of Health and Welfare Research Work for Treatment of Specific Diseases.
However, no study of the relation between dystrophy and calpain in retinal degenerative diseases has been found heretofore in the prior art.
OBJECTS OF THE INVENTION
The main object of the present invention is to investigate calpain which is tissue-specifically expressed in the retina of eye tissues, to isolate its gene, to determine the structure of a protein and to use them in studies of diseases in ophthalmologic field and in treatment and prevention of diseases in ophthalmologic field, in particular, retinal degenerative diseases.
This object as well as other objects and advantages of the present invention will become apparent to those skilled in the art with reference to the attached drawings.


REFERENCES:
patent: 0 717 110 (1996-06-01), None
patent: 96/16175 (1996-05-01), None
H Ma et al.,IOVS, “Cloning and Expression of mRNA for Calpain Lp82 from Rat Lens:Splice Variant of p94,” Feb. 1998, vol. 39, No. 2, pp. 454-460.*
H Ma et al., GenBank Accession No. U96367. Mar. 1998.*
Seikagaku (Biochemistry), vol. 65, pp. 537-552, 1993-together with an English translation of a pertinent part.
Jikken Igaku (Experimental Medicine), vol. 13, pp. 35-42, 1995-together with an English translation of a pertinent part.
Atarashii Ganak (Journal of the Eye), vol. 13, pp. 993-1001, 1996-together with an English translation of a pertinent part.
Atarashii Ganka (Journal of the Eye), vol. 12, pp. 239-250, 1995-together with an English translation of a pertinent part, and abstract of Nature 1990, Jan. 25:343 (related document).
H. Sorimachi et al., “Molecular Cloning of a Novel Mannalian Calcium-dependent Protease Distinct from Both m-and &mgr;-types”, The Journal of Biological Chemistry, vol. 264, No. 33, pp. 20106-20111, Nov. 25, 1989.
I. Richard et al., “Mutations in the Proteolytic Enzyme Calpain 3 Cause Limb-Girdle Muscular Dystrophy type 2A”, Cell, vol. 81, pp. 27-40, Apr. 7, 1995.
I. Richard et al., “Molecular Cloning of Mouse Canpe3, the Gene Associated with Limb-Girdle Muscular Dystrophy 2A in Human”, Mammalian Genome, vol. 7, pp. 377-379, 1996.
K. Wang et al., “Development and Therapeutic Potential of Calpain Inhibitors”, Advances in Pharmacology, vol. 37, pp. 117-152, 1997.
L. David et al., “Purification of Calpain II from Rat Lens and Determination of Endogenous Substrates”, Exp., Eye Research, vol. 42, pp. 227-238, 1986.
U.K. Laemmli, “Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4”, Nature, vol. 227, pp. 680-685, 1970.
H. Towbin et al., “Electrophoretic Transfer of Proteins from Polyacrylamide Gels to Nitrocellulose Sheets: Procedure and some Applications”, Proc. Natl. Acad. Sci., vol. 76, No. 9, pp. 4350-4354, Sep. 1979.
J. Biol. Chem., vol. 264, No. 33, 1989, Sorimachi H. et al., “Molecular cloning of a novel mammalian calcium-dependent prorease distinct from both m-and &mgr;-types”, pp. 20106-20111.
Cell, vol. 81, No. 5, 1995, Richard I. et al., “Mutations in the proteolytic enzyme calpain 3 cause limb-girdle muscular dystrophy type 2A”, pp. 27-40.
Mamm. Genome, vol. 7, No. 5, 1996, Richard I. et al., “Molecular cloning o mouse canp3, the gene associated with limb-girdle muscular dystrphy 2A in human”, pp. 377-379.
Biochim. Biophys. Acta, vol. 1261, No. 3, 1995, Sorimachi H. et al., “Identification of a third ubiquitous calpain species—chicken muscle expresses four distinct calpains”, pp. 381-393.

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